https://www.leica-microsystems.com/ Article tagged with en-US https://www.leica-microsystems.com/29672 Fluorescence Microscopy Widefield Microscopy Ovarian cancer is one of the most severe types of cancers that women can suffer from during their lifetimes. It is the cancer’s tendency for frequent relapses and drug resistance that leads to severity. High grade serous ovarian cancers are characterized by a special oxidative metabolism which has an influence on inflammation and drug resistance. TRAP1 is a chaperone which is involved in the regulation of mitochondrial respiration. https://www.leica-microsystems.com//science-lab/cholesterol-homeostasis-modulates-platinum-sensitivity-in-human-ovarian-cancer/ Tue, 02 Jun 2020 13:49:00 +0000 https://www.leica-microsystems.com/3815 Laser Microdissection Neuroscience Forensics This list presents current publications in the major application fields for laser microdissection and is updated monthly. https://www.leica-microsystems.com//science-lab/laser-microdissection-publication-list/ Wed, 27 May 2020 07:22:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/24935 Fluorescence Microscopy Live-Cell Imaging Widefield Microscopy To help you answer important scientific questions, THUNDER Imagers eliminate the out-of-focus blur that clouds the view of thick samples when using camera-based fluorescence microscopes. They achieve this using Computational Clearing our new opto-digital technology. The result is high-speed, high-quality imaging of a large diversity of three-dimensional samples, including model organisms, tissue sections, and 3D cell cultures. Take a look at these images to see how THUNDER Imagers are already helping researchers to reveal the finest structural details even deep within a sample. Once you have seen the haze-free image quality achieved with THUNDER Imagers, find out more on our product page. Learn how THUNDER Imagers combine Computational Clearing with the speed, fluorescence-signal sensitivity, and ease-of-use of widefield microscopes so you can decode 3D biology in real-time. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-thunder-imager/ Tue, 12 May 2020 11:20:00 +0000 https://www.leica-microsystems.com/29634 EM Sample Preparation CLEM In this article you can learn how to optimize the preparation quality of your samples by using the ion beam etching method with the EM TIC 3X ion beam milling machine. A short introduction of the instrument explains how the flexible setup of the EM TIC 3X gives you the opportunity to prepare samples for various investigation areas. https://www.leica-microsystems.com//science-lab/introduction-to-ion-beam-etching-with-the-em-tic-3x/ Mon, 11 May 2020 13:15:00 +0000 https://www.leica-microsystems.com/28338 Widefield Microscopy This webinar is designed to clarify crucial specifications that contribute to THUNDER Imagers' transformative visualization of 3D samples and improvements within a researcher's imaging-related workflow. https://www.leica-microsystems.com//science-lab/computational-clearing-enhance-3d-specimen-imaging/ Fri, 08 May 2020 12:56:00 +0000 https://www.leica-microsystems.com/26513 Quality Assurance Simultaneous visual and chemical analysis of steel non-metallic inclusions with a 2-methods-in-1 solution, using optical microscopy and laser induced breakdown spectroscopy (LIBS), is described in this report. For rapid, confident decisions during steel quality control or failure analysis, having both microstructural and composition data is important. A 2-in-1 solution can reduce the cost and time for accurate steel inclusion analysis. https://www.leica-microsystems.com//science-lab/visual-and-chemical-analysis-of-steel-microstructure-faster-rating-of-steel-quality/ Wed, 06 May 2020 12:38:00 +0000 PhD James DeRose, Dr. Kay Scheffler, PhD Konstantin Kartaschew, Mike Horz, Thomas Locherer https://www.leica-microsystems.com/29648 Confocal Microscopy In this article we explore several strategies you can take to improve the separation of fluorophores and increase the number of fluorescent probes you can distinguish in your sample. https://www.leica-microsystems.com//science-lab/explore-innovative-techniques-to-separate-fluorophores-with-overlapping-spectra/ Tue, 05 May 2020 09:59:00 +0000 PhD Liz Roquemore https://www.leica-microsystems.com/28596 Coherent Raman Scattering (CRS) Here we present a webinar on the emerging applications of Stimulated Raman Scattering Microscopy, covering the following topics: Quantitative spectroscopic imaging of label-free or chemically tagged specimens: From cell and tissue biology to medical research, materials science, and microfabrication; Probing 3D cellular model systems (organoids) and tissues for preclinical & translational research in cancer, neurodegenerative diseases, immunology, and digestive disorders; Multi-modal confocal nonlinear optical microscopy, combining CARS, SRS, Second-Harmonic Generation, and 2-Photon Fluorescence for advanced tissue-based diagnostics; Monitoring tissue penetration of pharmacological and cosmetic compounds https://www.leica-microsystems.com//science-lab/vibrational-spectroscopic-imaging-in-cells-tissues-and-model-organisms/ Wed, 29 Apr 2020 08:46:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/29614 Confocal Microscopy Leica Microsystems’ TauSense technology is a set of imaging modes based on fluorescence lifetime. Found at the core of the STELLARIS confocal platform, it will revolutionize your imaging experiments. Whatever your sample or staining procedure, fluorescence lifetime information is always there. Now TauSense gives you access to this additional information and expands the potential of your research with the possibilities provided by different TauSense modes. https://www.leica-microsystems.com//science-lab/add-an-extra-dimension-to-confocal-results-with-tausense/ Tue, 28 Apr 2020 15:13:00 +0000 https://www.leica-microsystems.com/29680 EM Sample Preparation This brochure presents and explains appropriate workflow solutions for the most frequently required sample preparation methods for material science samples. https://www.leica-microsystems.com//science-lab/workflow-solutions-for-industrial-research/ Tue, 28 Apr 2020 15:13:00 +0000 https://www.leica-microsystems.com/19946 Metallography Quality Assurance This webinar and report describe optimal microscopy solutions for rating steel quality in terms of non-metallic inclusions and reviews the various international and regional standards concerning rigorous quality assessment methods, e.g., EN 10247, ASTM E45, DIN 50602, and ISO 4967. https://www.leica-microsystems.com//science-lab/rate-the-quality-of-your-steel-free-webinar-and-report/ Tue, 28 Apr 2020 12:19:00 +0000 Dionis Diez, PhD James DeRose, Thomas Locherer https://www.leica-microsystems.com/28312 Metallography Learn how to get spatially resolved information about the chemical composition of elements in seconds - quickly and easily. https://www.leica-microsystems.com//science-lab/simplify-and-speed-up-element-analysis-at-the-micron-range/ Wed, 22 Apr 2020 15:43:00 +0000 https://www.leica-microsystems.com/28460 Confocal Microscopy The vast majority of imaging experiments measure fluorescence intensity but there is another key property of fluorescence—its lifetime, which is information that comes “for free” with every experiment. In this article we explore what lifetime imaging is, and how it can improve quality and add depth to your applications. https://www.leica-microsystems.com//science-lab/how-to-uncover-hidden-dimensions-in-research-with-lifetime-imaging/ Wed, 15 Apr 2020 11:27:00 +0000 PhD Liz Roquemore https://www.leica-microsystems.com/28402 Surgical Microscopy The tutorial features the Leica ARveo digital Augmented Reality microscope for complex neurosurgery. The procedure also applies to the Leica M530 OHX, OH6, OH5 and OH4. https://www.leica-microsystems.com//science-lab/how-to-drape-an-overhead-surgical-microscope/ Mon, 06 Apr 2020 13:14:00 +0000 https://www.leica-microsystems.com/28397 Surgical Microscopy Before performing surgical procedures, it is important to drape the surgical microscope to ensure sterile working conditions. At Leica, we are committed to helping you with your surgical practice. In the below video, discover a step-by-step tutorial on draping your surgical microscope. The tutorial features the Leica PROvido multidisciplinary surgical microscope. The procedure also applies to the Leica M525 F50 microscope. https://www.leica-microsystems.com//science-lab/how-to-drape-a-surgical-microscope/ Mon, 06 Apr 2020 13:07:00 +0000 https://www.leica-microsystems.com/28354 Basics in Microscopy Due to the current coronavirus pandemic, there are a lot of questions regarding decontamination methods of microscopes for safe usage. This informative article summarizes general decontamination methods and recommendations without being meant to replace a scientists’ expertise nor medical or professional advice. https://www.leica-microsystems.com//science-lab/how-to-sanitize-a-microscope/ Fri, 03 Apr 2020 09:20:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/28381 Surgical Microscopy Dr. Salam Abu Arqub, from the Smile Engineer Dental Center in Amman, Jordan, has been using Leica dental microscopes for three years for all procedures performed at the clinic. He shared his experience at the AEEDC Dental Congress in Dubai. https://www.leica-microsystems.com//science-lab/overcoming-complexities-in-microdentistry/ Wed, 01 Apr 2020 15:43:00 +0000 https://www.leica-microsystems.com/28376 Surgical Microscopy Microscopes do not only provide better visualization during dental surgery. They also help ensure correct posture to avoid back pain and neck injuries. Dr. Iyad Ghoneim, from the Safad Dental Center in Riyadh, Saudi Arabia, shared his experience working with the Leica PROvido surgical microscope during a presentation at the AEEDC Dental Congress in Dubai. https://www.leica-microsystems.com//science-lab/minimally-invasive-dentistry-visualization-posture/ Wed, 01 Apr 2020 13:48:00 +0000 https://www.leica-microsystems.com/27800 Widefield Microscopy This webinar will showcase the versatility and performance of THUNDER Imagers in many different life science applications: from counting nuclei in retina sections and RNA molecules in cancer tissue sections to monitoring calcium waves in Arabidopsis seedlings and much more. https://www.leica-microsystems.com//science-lab/thunder-imagers-high-performance-versatility-and-ease-of-use-for-your-everyday-imaging-workflows/ Wed, 25 Mar 2020 12:56:00 +0000 https://www.leica-microsystems.com/28307 Confocal Microscopy Autofluorescence can significantly reduce what you can see in a confocal experiment. This article explores causes of autofluorescence as well as different ways to remove it, from simple media fixes to powerful technological solutions. Removing this background is essential to correctly interpreting small differences in the signals present within your sample. Find out how in this article. https://www.leica-microsystems.com//science-lab/learn-how-to-remove-autofluorescence-from-your-confocal-images/ Mon, 23 Mar 2020 09:58:00 +0000 PhD Ben Libberton https://www.leica-microsystems.com/28066 Widefield Microscopy Damaged nerve regeneration was investigated using mouse spinal cord sections treated with compounds that counter axon growth inhibitor (AGI) proteins. The sections were screened to find active and non-active axons using widefield and THUNDER imaging technology. Results indicated a better discrimination between active and non-active axons in THUNDER images. https://www.leica-microsystems.com//science-lab/evaluating-axon-regeneration-after-brain-or-spine-trauma-of-mice/ Tue, 17 Mar 2020 11:55:00 +0000 PhD James DeRose, Robert Fasulka https://www.leica-microsystems.com/28278 Surgical Microscopy Prof. Kei Shinoda invited four surgeons, all experts in optical performance and microscope systems, to talk about the Proveo 8 ophthalmic microscope from Leica Microsystems. The group discussed topics like the future of ophthalmic surgery and the role that microscopes will play in it. https://www.leica-microsystems.com//science-lab/ophthalmic-surgical-panel-discussion-summary/ Mon, 16 Mar 2020 13:59:00 +0000 Prof. Kei Shinoda, Dr. Toshiaki Kurakazu, Dr. Tatsuhiro Nakamura, Dr. Shunsuke Osawa, Dr. Masayuki Shibuya https://www.leica-microsystems.com/27942 Fluorescence Microscopy Widefield Microscopy Magali Mondin, an engineer at the BIC (Bordeaux Imaging Center) in France, describes her experience using a THUNDER Imager in this interview. She had the opportunity to test extensively the THUNDER Imager 3D Tissue in her lab for several weeks on a wide variety of specimens. We met with her after the testing was finished to discuss the results she obtained and her experience working with the THUNDER technology. The BIC offers resources for photonic and electronic imaging, mainly in the life, health, and plant sciences. It is a core facility in France with a staff of 12 highly skilled engineers. The BIC has advanced bio-imaging techniques for fixed- and live-cell imaging, such as video microscopy, confocal microscopy, multiphoton microscopy, transmission electron microscopy, and scanning electron microscopy. https://www.leica-microsystems.com//science-lab/interview-with-magali-mondin-on-thunder-imagers/ Mon, 09 Mar 2020 12:43:00 +0000 Engineer Magali Mondin https://www.leica-microsystems.com/28027 Surgical Microscopy Building on a decade of leadership in fluorescence imaging technology, GLOW800 AR fluorescence is the first of many modalities based on the proprietary GLOW AR platform. The sophisticated imaging sensors and algorithms of GLOW AR acquire, optimize, and combine multiple spectral bands of light. The result is natural coloring of tissue anatomy and accurate representation of fluorescence intensity. https://www.leica-microsystems.com//science-lab/galleries/augmented-reality-ar-fluorescence-image-gallery/ Thu, 05 Mar 2020 10:28:00 +0000 MD Robert Ibe https://www.leica-microsystems.com/27966 Surgical Microscopy Managing unruptured intracranial aneurysms is complex and usually involves one or a combination of three options – conservative, endovascular, or surgical treatment. During surgical treatment the ability to simultaneously visualize and assess the anatomical structures and blood flow is one of the biggest challenges of neurovascular surgery. It is even more challenging to view this in real time. Aneurysms, like all other vascular malformations, are delicate structures with the potential to rapidly deteriorate. Having imaging techniques that provide adequate visualization and assessment of both blood flow and the related anatomical structures enhances decision making. https://www.leica-microsystems.com//science-lab/unruptured-intracranial-aneurysms/ Fri, 21 Feb 2020 15:57:00 +0000 Eugenia Petoukhov, MD Robert Ibe https://www.leica-microsystems.com/19049 Multiphoton Microscopy Live-Cell Imaging Confocal Microscopy Fluorescence Microscopy Neuroscience Multiphoton Microscopy is an advanced technique for imaging thick samples. Applications range from the visualization of the complex architecture of the whole brain to the study of tumor development and metastasis or the responses of the immune system in living animals. On this regularly updated reference list you can find selected publications on reseach using multiphoton microscopy. https://www.leica-microsystems.com//science-lab/multiphoton-microscopy-publication-list/ Thu, 20 Feb 2020 08:47:00 +0000 https://www.leica-microsystems.com/27674 Coherent Raman Scattering (CRS) Stimulated Raman Scattering (SRS) is an emerging microscopy technique that images the vibrations of chemical bonds within a specimen. It allows researchers to probe the endogenous chemistry or biochemistry of their specimen without the need for exogenous labels. https://www.leica-microsystems.com//science-lab/how-to-prepare-samples-for-stimulated-raman-scattering/ Tue, 11 Feb 2020 08:59:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/12775 Quality Assurance In the automobile industry, even the smallest particles on single components can cause malfunctioning. Fuel, lubricant and urea filter systems are particularly vulnerable. Component cleanliness has therefore become a central quality characteristic in the modern manufacturing process, especially when fitting components of various suppliers. https://www.leica-microsystems.com//science-lab/key-factors-for-efficient-cleanliness-analysis/ Fri, 07 Feb 2020 12:27:00 +0000 PhD James DeRose, Dr. Kay Scheffler, PhD David R. Barbero https://www.leica-microsystems.com/27780 Surgical Microscopy This case study from Prof. Dr. Feres Chaddad talks about the treatment of unruptured MCA (middle cerebral artery) and PCOM (posterior communicating artery) aneurysms with microsurgical clipping. It illustrates how the Augmented Reality Fluorescence GLOW800 can help surgeons before and after clipping with one augmented, real-time view of the cerebral anatomy, augmented by real-time vascular flow. https://www.leica-microsystems.com//science-lab/glow800-augmented-reality-fluorescence-in-aneurysm-treatment/ Mon, 03 Feb 2020 12:46:00 +0000 Prof Dr. Feres Chaddad, MD Robert Ibe https://www.leica-microsystems.com/27634 Quality Assurance Polarized light microscopy (also known as polarizing microscopy) is an important method used in different fields, including research and quality assurance. It goes beyond just producing images at high magnification and resolution, something typically done with microscopes using ordinary optics. By examining the form, structure, color, birefringence, and further optical properties, additional information about the sample structure, optical properties, and composition can be obtained. https://www.leica-microsystems.com//science-lab/galleries/polarizing-microscope-image-gallery/ Wed, 29 Jan 2020 10:58:00 +0000 https://www.leica-microsystems.com/11172 Metallography Quality Assurance This article gives an overview of metallography and metallic alloy characterization. Different microscopy techniques are used to study the alloy microstructure, i.e., microscale structure of grains, phases, inclusions, etc. Metallography developed from the need to understand the influence of alloy microstructure on macroscopic properties. The knowledge obtained is exploited for the design, development, and manufacture of alloy materials. https://www.leica-microsystems.com//science-lab/metallography-an-introduction/ Wed, 08 Jan 2020 12:43:00 +0000 Dionis Diez, PhD James DeRose https://www.leica-microsystems.com/6654 Multiphoton Microscopy Basics of multiphoton microscopy. This tutorial explains the principles of multiphoton microscopy for deep tissue imaging. Multiphoton microscopy uses excitation wavelengths in the infrared taking advantage of the reduced scattering of longer wavelengths. This makes multiphoton imaging the perfect tool for deep tissue imaging in thick sections and living animals. https://www.leica-microsystems.com//science-lab/principles-of-multiphoton-microscopy-for-deep-tissue-imaging/ Mon, 16 Dec 2019 11:42:00 +0000 Dr. Andrea Mülter https://www.leica-microsystems.com/4592 Coherent Raman Scattering (CRS) CARS (Coherent Anti-Stokes Raman Scattering) microscopy is a dye-free method which images structures by displaying the characteristic intrinsic vibrational contrast of their molecules. The crucial advantage of this method is that the sample remains almost unaffected. This tutorial explains the molecular basics on Coherent Anti-Stokes Raman Scattering. https://www.leica-microsystems.com//science-lab/step-by-step-guide-to-the-molecular-basics-of-cars-microscopy/ Mon, 16 Dec 2019 11:38:00 +0000 Dr. Stefanie Degenhartt https://www.leica-microsystems.com/5098 Stereo Microscopy Ergonomics Basics in Microscopy Quality Assurance Stereo microscopes are often nicknamed the “workhorse” of the lab or production site. Users spend many hours looking through the eyepieces inspecting, observing, documenting, or dissecting samples. Careful assessment of the relevant applications for which the stereo microscope is needed is key to long-lasting, satisfactory use. https://www.leica-microsystems.com//science-lab/factors-to-consider-when-selecting-a-stereo-microscope/ Wed, 11 Dec 2019 07:00:00 +0000 B.Sc. Elisabeth Ippel, PhD James DeRose, Daniel Goeggel https://www.leica-microsystems.com/27386 Super-Resolution The endoplasmic reticulum (ER) is a continuous membrane organelle in charge of protein synthesis, lipid synthesis and detoxification. The ER structure is described in terms of smooth peripheral tubules and rough ER sheets. The family of proteins responsible for maintaining the sheet or tubule architecture include the cytoskeleton-linking membrane protein 63 (CLIMP-63) and reticulon (RTN), respectively. A detailed map of the nanodomain distribution remains challenging due to the size of the structures and their highly dynamic nature. To give an idea, the ER sheet thickness and ER tubule diameter are in the 30-100 nm range, well below the diffraction limit of confocal microscopy. https://www.leica-microsystems.com//science-lab/new-light-shed-on-the-nanodomain-organization-of-the-endoplasmic-reticulum-er/ Mon, 02 Dec 2019 10:59:00 +0000 https://www.leica-microsystems.com/27272 Coherent Raman Scattering (CRS) Despite decades of research, the molecular mechanisms underlying some of the most severe neurodegenerative diseases, such as Alzheimer’s or Parkinson’s, remain poorly understood. The progression of these diseases often correlates with the appearance of abnormal aggregations of proteins and lipids in affected brain tissues. There is hope that detailed investigations of these pathological structures might yield new insights into the disease mechanisms. Hence, there is a strong demand for advanced analytical techniques that can probe these structures in intact tissues. Here, we show that stimulated Raman scattering (SRS) microscopy can provide label-free, high-resolution, chemically specific characterization of tissue pathologies associated with neurodegenerative diseases. https://www.leica-microsystems.com//science-lab/stimulated-raman-scattering-microscopy-probes-neurodegenerative-disease/ Wed, 20 Nov 2019 11:32:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/27289 Metallography Watch this webinar to learn more about obtaining reliable and accurate metallographic results with Leica microscopes in terms of efficient analysis of non-metallic inclusions in steel as well as of alloy grains and phases. Develop better, cleaner products with speed and reliability using flexible and affordable metallographic microscopy solutions from Leica Microsystems. The solutions help you reduce costs by efficiently providing statistically meaningful data for the continuous improvement of your production workflow. https://www.leica-microsystems.com//science-lab/metallographic-solutions-from-leica-microsystems/ Mon, 11 Nov 2019 12:01:00 +0000 https://www.leica-microsystems.com/26832 Quantitative Imaging This video shows the presentation of Dr. Kohki Okabe given at the 9th Congress of the Federation of Asian and Oceanian Physiological Society (FAOPS) held in Kobe, Japan on 30 March 2019. Dr. Okabe is an Assistant Professor and JST-PRESTO Researcher with the Laboratory of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo. https://www.leica-microsystems.com//science-lab/imaging-intracellular-temperature-using-fluorescence-lifetime-imaging-microscopy-flim/ Thu, 07 Nov 2019 09:10:00 +0000 PhD Kohki Okabe, Dr. Giulia Ossato https://www.leica-microsystems.com/26359 Laser Microdissection This study focused on tissue-specific metabolite profiling and comparison of various parts of bolting and unbolting Peucedanum praeruptorum Dunn (Qianhu in Chinese). The qualitative and quantitative results were used to empirically clarify the suitability of Qianhu for medicinal purposes. https://www.leica-microsystems.com//science-lab/tissue-specific-metabolite-profiling-on-the-different-parts-of-bolting-and-unbolting-peucedanum-praeruptorum-dunn-qianhu/ Wed, 06 Nov 2019 09:54:00 +0000 Eugenia Petoukhov https://www.leica-microsystems.com/11641 Surface Metrology Quality Assurance This report briefly discusses several important metrology techniques and standard definitions commonly used to assess the topography of surfaces, also known as surface texture or surface finish. With the advent of nanotechnology, thin coatings, and the miniaturization of circuits and devices, surface metrology has become an extremely important field of science and engineering. https://www.leica-microsystems.com//science-lab/brief-introduction-to-surface-metrology/ Wed, 06 Nov 2019 08:27:00 +0000 PhD James DeRose, M.S. Albert S. Laforet, PhD David R. Barbero https://www.leica-microsystems.com/26925 Widefield Microscopy Differentiated living beings such as humans, but also a fruit fly or a plant, possess not only the differentiated cells which form specific tissues, but also those cells whose fate is not yet (or only partially) fixed. Such cells are called stem cells. Stem cells serve to proliferate new cells for the specific tasks, but at the same time always produce stem cells, which in turn can supply new specific cells when needed later. https://www.leica-microsystems.com//science-lab/drosophila-testis-niche-stem-cells-three-color-computational-clearing/ Mon, 04 Nov 2019 11:42:00 +0000 Linda B. Morgan, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/26722 Multiphoton Microscopy Dr. Thomas Mathivet, working at the Cardiovascular Institute in Paris, France, was interviewed by Dr. Luis Alvarez, Product Application Manager, during Leica Microsystems Meets Science 2019 in Mannheim, Germany. He discusses his research on the cardiovascular system in the brain. The goal is a better understanding of specific disease conditions, such as brain tumors. Dr. Mathivet’s team performs real-time imaging of the brain in live animals and the results play a significant role in the development of new intervention strategies for these pathologies. The SP8 DIVE spectrally tunable multiphoton microscope has changed the approach of his team to imaging. The need to visualize simultaneously with 5 to 6 different channels, not possible with traditional systems, is now met. Dr. Mathivet has found that the new imaging capabilities of the SP8 DIVE enable him to see more, including label-free visualization of collagen to study the brain’s structural aspects. https://www.leica-microsystems.com//science-lab/chat-with-dr-thomas-mathivet-about-in-vivo-cardiovascular-imaging-of-the-brain/ Wed, 30 Oct 2019 11:34:00 +0000 PhD Thomas Mathivet, Dr. Luis Alvarez https://www.leica-microsystems.com/26903 Confocal Microscopy This work reports new interesting findings concerning porous media infiltration which have direct implications for the fields of energy (oil recovery) and environment (CO2 sequestration and aquifer remediation). https://www.leica-microsystems.com//science-lab/pore-scale-visualization-of-drainage-in-3d-porous-media-by-confocal-microscopy/ Mon, 28 Oct 2019 07:07:00 +0000 D.Sc. Débora F. do Nascimento, M.Sc. Ronaldo Vimieiro Junior, PhD David R. Barbero, PhD Marcio Carvalho https://www.leica-microsystems.com/26877 EM Sample Preparation Neuroscience Intracellular dynamic events, such as protein trafficking, recycling, and degradation as well as signal transduction, to name a few, can be studied using live-cell imaging techniques. However, conventional light microscopes have limited resolution which is not sufficient to visualize cellular processes that occur at the nanometer scale in their cellular context. Electron microscopy is widely employed to visualize molecular complexes, cellular organelles, tissues and whole organisms at the ultrastructural level. https://www.leica-microsystems.com//science-lab/ultrastructural-preservation-and-improved-visualization-of-membranes-in-primary-bovine-chromaffin-cells/ Mon, 21 Oct 2019 13:45:00 +0000 MSc Gianvito Arpino, MD, PhD Ling-Gang Wu, PhD Christopher Bleck https://www.leica-microsystems.com/26677 Quantitative Imaging SP8 FALCON (FAst Lifetime CONtrast) is a fast and completely integrated fluorescence lifetime imaging microscopy (FLIM) confocal platform. SP8 FALCON delivers video-rate FLIM with pixel-by-pixel quantification, thanks to a novel concept for measuring fluorescence lifetimes built on fast electronics and sensitive spectral hybrid detectors. Photon arrival times are recorded at count rates typical for standard confocal imaging. The system has ultra-short dead time and powerful built-in algorithms for data acquisition and analysis. The deep integration of FLIM into the confocal platform provides easy access to complex FLIM experiments. https://www.leica-microsystems.com//science-lab/a-novel-concept-in-fluorescence-lifetime-imaging-enabling-video-rate-confocal-flim/ Thu, 17 Oct 2019 09:37:00 +0000 Dr. Luis Alvarez, Bernd Widzgowski, Bram van den Broek, Dr. Giulia Ossato, PhD Kees Jalink, Dr. Lioba Kuschel, Dr. Julia Roberti, Frank Hecht https://www.leica-microsystems.com/20212 EM Sample Preparation Metallography Quality Assurance Stereo Microscopy The investigation of macroscale defects in aluminum (Al) alloys with a rapid 3-dimensional (3D) imaging approach is described in this report. Aluminum (Al) alloys play an important role in the production of aircraft and vehicles, as well as products in other industries. Defects present in the Al alloy used for the production of aircraft, vehicles, or other products can have a significant effect on their quality, performance, and lifetime. https://www.leica-microsystems.com//science-lab/3-dimensional-imaging-of-macroscopic-defects-in-aluminum-alloys/ Thu, 17 Oct 2019 07:00:00 +0000 PhD Wolfgang Grünewald, PhD James DeRose https://www.leica-microsystems.com/26685 EM Sample Preparation The potential of cryogenic broad ion beam milling used in combination with scanning electron microscopy (cryo-BIB-SEM) for imaging and analyzing the microstructure of cryogenically stabilized soft polymers is assessed in this report. Results from the examination of delicate, natural polymers, such as tomato skin and wood, with cryo-BIB-SEM are shown. The polymer surface morphology and various microstructural features were analyzed. https://www.leica-microsystems.com//science-lab/studying-the-microstructure-of-natural-polymers-in-fine-detail/ Wed, 16 Oct 2019 10:30:00 +0000 PhD Joyce Schmatz, PhD James DeRose, PhD David R. Barbero, M.Sc. Martin Nopens https://www.leica-microsystems.com/4421 Coherent Raman Scattering (CRS) CRS (Coherent Raman Scattering) microscopy is an umbrella term for label-free methods that image biological structures by exploiting the characteristic, intrinsic vibrational contrast of their molecules. The two most important CRS techniques are Coherent Anti-Stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS). The biochemical image contrast of CRS is in many ways complementary to the molecular contrast obtained in fluorescence microscopy. A second crucial advantage of these methods is that they preserve the specimen/sample in a near pristine state. This reference list presents current and basic papers on CRS microscopy. https://www.leica-microsystems.com//science-lab/cars-publication-list/ Tue, 08 Oct 2019 22:00:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/14345 EM Sample Preparation When studying samples, to visualize their fine structure with nanometer scale resolution, most often electron microscopy is used. There are 2 types: scanning electron microscopy (SEM) which images the sample surface or transmission electron microscopy (TEM) which requires a very thin, electron-transparent sample. Thus, to image the fine structure inside a sample using electron microscopy, the solution is to make very thin sections of it. https://www.leica-microsystems.com//science-lab/brief-introduction-to-ultramicrotomy/ Tue, 17 Sep 2019 09:06:00 +0000 Robert Ranner, PhD James DeRose https://www.leica-microsystems.com/6282 TIRF Microscopy This monthly updated references list presents current papers using Leica AM TIRF in the major application fields for TIRF microscopy. https://www.leica-microsystems.com//science-lab/tirf-publication-list/ Tue, 03 Sep 2019 12:50:00 +0000 https://www.leica-microsystems.com/27911 CLEM Leica Microsystems and Thermo Fisher Scientific have collaborated to create a fully integrated cryo-tomography workflow that responds to these research needs: Reveal cellular mechanisms at subnanometer resolution in 3D applying the fully integrated cryo electron tomography workflow https://www.leica-microsystems.com//science-lab/improve-cryo-electron-tomography-workflow/ Tue, 03 Sep 2019 09:50:00 +0000 https://www.leica-microsystems.com/26294 Quantitative Imaging Prof. Ammasi Periasamy, Director, Keck Center for Cellular Imaging, University of Virginia, was interviewed by Dr. Giulia Ossato, Product Manager functional imaging, during Leica Microsystems Meets Science 2019 in Mannheim, Germany. They had an inspiring chat about fluorescence lifetime imaging microscopy (FLIM). The technological development of FLIM and its relevance for functional imaging in terms of studying redox states was discussed. Prof. Periasamy also talked about his research on metabolism and dysfunction in live cells. He stated that one of the central developments in the last few decades is the application of lifetime contrast to gather information about molecular interactions and function. https://www.leica-microsystems.com//science-lab/development-of-fluorescence-lifetime-imaging-microscopy-flim-and-its-relevance-for-functional-imaging/ Wed, 28 Aug 2019 10:13:00 +0000 Professor Ammasi Periasamy, Dr. Giulia Ossato https://www.leica-microsystems.com/26346 Multiphoton Microscopy Confocal Microscopy The renal corpuscle of the kidney comprises a glomerular vasculature embraced by podocytes and supported by mesangial myofibroblasts. Their combined actions are essential for the formation of an acellular plasma filtrate that passes through the podocyte-generated slit diaphragm into the nephron. Mutations in podocyte-expressed genes lead to chronic disease. An enhanced understanding of podocyte development and function to create biologically relevant in vitro podocyte models is a clinical imperative. Our team studied human podocyte development using scRNA-seq technology and established a framework to unbiasedly evaluate human embryonic stem cells (hESC)-derived podocytes. Confocal imaging served as a powerful tool for us to capture compelling immunofluorescent and fluorescent in situ hybridization validations of transcriptomic findings. https://www.leica-microsystems.com//science-lab/multiphoton-imaging-bolsters-findings-on-in-vitro-differentiation-of-pluripotent-stem-cell-derived-podocyte/ Mon, 26 Aug 2019 11:38:00 +0000 Tracy Tran https://www.leica-microsystems.com/25109 Surgical Microscopy Prof. Sleiman talks about the most important features for good imaging during his dental procedures using a Leica dental microscope. A large depth of field and a powerful illumination are very important for instance to correctly identify details, colors, and shapes inside an access cavity. https://www.leica-microsystems.com//science-lab/how-to-improve-clinical-practice-with-dental-microscope-solutions-from-leica-microsystems/ Mon, 19 Aug 2019 13:32:00 +0000 Prof. Philippe W. Sleiman https://www.leica-microsystems.com/26320 Live-Cell Imaging TIRF Microscopy Plants, Bacteria, and Fungi possess a rigid cell wall that protects the cell and gives it shape. Animal cells, such as mammalian cells, have no outer wall, which exposes their plasma membrane to the environment. Still, they have a distinct shape, can easily alter their shape, and even change their shape to move around. This is possible thanks to a flexible “inner wall” that is composed of the actin cortex and the cytoskeleton. https://www.leica-microsystems.com//science-lab/where-to-go-cellular-migration-requires-coordinated-transitions-of-actin-cortex/ Mon, 19 Aug 2019 13:27:00 +0000 PhD Olga Davydenko, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/26318 Neuroscience Confocal Microscopy Cellular motility is based on motor-proteins that can bind to filamentous scaffold proteins and – under consumption of ATP – can “crawl” on these filaments. This note is about proteins connected to microtubules, one of the filamentous structures that compose the cytoskeleton. Microtubules are hollow tubes of ca 25nm, composed of tubulin-heterodimers. The proteins are polymerized in a directed fashion, allowing to differentiate a plus-end and a minus-end of the fiber. Another important scaffold component that is also involved in movements, are actin fibers that cooperate with myosin as a motor-protein. The best known movement involving the actin-myosin system is muscular contraction. https://www.leica-microsystems.com//science-lab/cellular-motility-microtubules-motor-proteins-and-tau-proteins/ Mon, 12 Aug 2019 12:34:00 +0000 PhD Jen-Yi Lee, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/20057 Quality Assurance This report explains how a 2-in-1 materials analysis solution, combining optical microscopy and chemical analysis by laser induced breakdown spectroscopy (LIBS), leads to an overall more efficient and cost-effective cleanliness workflow. https://www.leica-microsystems.com//science-lab/cleanliness-analysis-with-a-2-methods-in-1-solution/ Wed, 07 Aug 2019 22:00:00 +0000 PhD James DeRose, Dr. Kay Scheffler https://www.leica-microsystems.com/19781 Metallography Metallic alloys, such as steel and aluminum, have an important role in a variety of industries, including automotive and transportation. In this report, the importance of grain size analysis for alloy characterization is reviewed, along with a practical, flexible microscopy solution using a high-performance software for easy user operation and image analysis. https://www.leica-microsystems.com//science-lab/how-to-adapt-grain-size-analysis-of-metallic-alloys-to-your-needs/ Wed, 31 Jul 2019 23:00:00 +0000 Dionis Diez, PhD James DeRose https://www.leica-microsystems.com/25143 Quantitative Imaging In her latest studies, Dr. Mukta Deobagkar-Lele investigated the role of ZIP7, a zinc transporter protein, in modulating B cell activity and its effects on human immunity. During an interview with Dr. Luis Alvarez from Leica Microsystems, she describes the key findings of these studies published in her recent article in Nature Immunology. She also discusses her introduction to lifetime imaging with the SP8 FALCON and how this modality helps her explore B-cell biology and related immunodeficiencies. https://www.leica-microsystems.com//science-lab/how-fretflim-biosensors-and-lifetime-imaging-helped-identify-the-importance-of-zinc-for-human-b-cell-development/ Tue, 30 Jul 2019 09:10:00 +0000 PhD Mukta Deobagkar, Dr. Luis Alvarez https://www.leica-microsystems.com/26267 Fluorescence Microscopy Stereo Microscopy Autofluorescence can be a very practical tool for the visualization of growth rings in wood, especially when these structures are hard to distinguish, like in tropical trees. Tree rings are largely used for the investigation of growth rate, forest dynamics and management, paleoclimate and climate change, among other applications. https://www.leica-microsystems.com//science-lab/improved-tree-ring-visualization-using-autofluorescence/ Tue, 23 Jul 2019 16:00:00 +0000 Ph.D. Piero Bagnaresi, PhD James DeRose https://www.leica-microsystems.com/26264 Super-Resolution This guide is intended to help users optimize sample preparation for stimulated emission depletion (STED) nanoscopy, specifically when using the TCS SP8 STED 3X nanoscope from Leica Microsystems. It gives an overview of fluorescent labels used for single color STED imaging and a ranking of their performance. Fluorescent label combinations for dual and triple color STED imaging that minimize cross-talk during detection are recommended. There is a discussion of considerations for immunofluorescence labeling and a detailed protocol to obtain high quality images, with a high signal/noise (S/N) ratio, of interesting structures in a specimen. Important details for sample mounting and substrates that enable optimal imaging, minimizing aberrations and autofluorescence due to the mounting medium, are reviewed. Finally, for STED imaging of live-cells, the most appropriate fluorescent labels are mentioned, both fluorescent proteins (FPs) and organic fluorophores which give the best performance. https://www.leica-microsystems.com//science-lab/the-guide-to-sted-sample-preparation/ Mon, 22 Jul 2019 17:00:00 +0000 https://www.leica-microsystems.com/25048 Education Widefield Microscopy Digital Microscopy As teachers, you know your big challenge is to catch and keep the students’ attention and the best chance for this is by making the environment interactive. In the case of the Microscopy Classroom, we can create an interactive environment using digital imaging components such as cameras, apps, and software. This webinar will familiarize you with the various methods you and your students can use to connect to a microscope camera and increase the learning time while enhancing the learning experience. https://www.leica-microsystems.com//science-lab/digital-classroom-options/ Tue, 09 Jul 2019 22:00:00 +0000 Vince Vaccarelli https://www.leica-microsystems.com/25096 Widefield Microscopy More than 60% of all diagnosed cases of dementia are attributed to Alzheimer’s disease. Typical of this disease are histological alterations in the brain tissue. So far, there is no cure for this disease. A few treatments try to slow down the fatal path or try to relieve the patients from symptoms. The laboratory of Dr. Mehrdad Shamloo at Stanford University studies pathological brain function with the goal of contributing to the discovery of novel therapeutics for Alzheimer’s disease. They use a mouse model of this disease to study the role of inflammation in Alzheimer’s disease progression. This requires imaging of thick uncleared brain tissues. https://www.leica-microsystems.com//science-lab/alzheimer-plaques-fast-visualization-in-thick-sections/ Thu, 27 Jun 2019 10:50:00 +0000 PhD Olga Davydenko, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/25094 Quantitative Imaging To perform quantitative studies of membrane bound G protein-coupled receptors (GPCRs) with high-performance microscopy, the proteins must first be labeled with bright and photostable dyes. The labeling method should be minimally invasive to avoid perturbing the GPCR function and result in quantitative yields to allow stoichiometric data analysis. For the precise quantification of dye yields, a method based on fluorescence fluctuation microscopy (molecular brightness) has been developed. The method can extract the number of labeling sites at the single-cell level. Because the reported technique is performed using a commercially available microscope, the approach can be used for the general study of membrane proteins with fluorescence microscopy. https://www.leica-microsystems.com//science-lab/single-residue-bioorthogonal-labeling-of-g-protein-coupled-receptors-gpcrs-in-live-cells-for-quantitative-analysis/ Wed, 26 Jun 2019 10:03:00 +0000 PhD James DeRose, PhD Paolo Annibale https://www.leica-microsystems.com/25036 EM Sample Preparation Get an insight in the working methods of the laboratory and learn about the advantages of Cryo SEM investigation in EM Sample Preparation. Find out how high pressure freezing, freeze fracturing and cryo transfer add to the Cryo SEM workflow and how the Leica portfolio ensures the compatibility between these different steps. https://www.leica-microsystems.com//science-lab/expert-knowledge-on-high-pressure-freezing-and-freeze-fracturing-in-the-cryo-sem-workflow/ Thu, 23 May 2019 22:00:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/25016 Quality Assurance Metallography Visual and chemical analysis of contamination on materials with a 2-methods-in-1 solution leading to an efficient, more complete analysis workflow is described in this report. A 2-in-1 solution, combining optical microscopy and laser induced breakdown spectroscopy (LIBS), in addition to simultaneous visual and chemical inspection, can be used to quickly remove contamination and inspect the underlying base material. For inspection of components or parts, such as soldering and leads on printed circuit boards (electronics) or metal panels on vehicles (automotive and transport), understanding the effects of contamination are important. https://www.leica-microsystems.com//science-lab/fast-visual-and-chemical-analysis-of-contamination-and-underlying-layers/ Wed, 22 May 2019 22:00:00 +0000 PhD James DeRose, Mike Horz, Dr. Kay Scheffler https://www.leica-microsystems.com/25023 Confocal Microscopy The marvelous complexity of life is perhaps the greatest source of fascination within the biological sciences. Among the most complex subjects within biology is the development of multicellular life. Differentiating from a single cell, multicellular organisms generate remarkably intricate and complex patterns and forms. It seems, therefore, almost counterintuitive that the long history of studying genetic regulation in ontogenesis is so closely and intrinsically entangled with research on an otherwise unremarkable organism; an organism more commonly recognized as a nuisance, the fruit fly Drosophila melanogaster. https://www.leica-microsystems.com//science-lab/visualize-developmental-patterns-in-drosophila/ Tue, 21 May 2019 22:00:00 +0000 Ryan E. Robinson, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/25022 EM Sample Preparation Nanoscale ultrastructural information is typically obtained by means of static imaging of a fixed and processed specimen. However, this is only a snapshot of one moment within a dynamic system in which structures are constantly changing. Exploring specific time points of a dynamic process is therefore a major challenge. Exploring a process at the nanoscale through optogenetics or electrical field stimulation in combination with timed millisecond precision vitrification is a promising technology to overcome this challenge. In the first part of a series of application notes the practical considerations of stimulation-assisted vitrification are discussed. https://www.leica-microsystems.com//science-lab/bridging-structure-and-dynamics-at-the-nanoscale-through-optogenetics-and-electrical-stimulation/ Mon, 20 May 2019 22:00:00 +0000 Dr. Andres Kaech, PhD Frédéric Leroux https://www.leica-microsystems.com/24965 Digital Microscopy Stereo Microscopy Quality Assurance Watch this video to see how you can rapidly record sharp optical microscope images of samples with a large height variation. This is done with the optional Extended Depth of Focus (EDOF) function of the LAS X software from Leica Microsystems. Acquisition of EDOF images of a printed circuit board, recorded from low to high magnification using a Leica microscope, are shown as an example. The operation of EDOF by acquiring a z-stack of images in 3 modes is demonstrated. https://www.leica-microsystems.com//science-lab/how-to-create-edof-extended-depth-of-focus-images/ Wed, 08 May 2019 22:00:00 +0000 Clinton Smith, Mike Horz https://www.leica-microsystems.com/24967 Digital Microscopy Surface Metrology Quality Assurance Watch this video to learn how you can save time when doing topographic analysis using the Leica Map software. The software allows you to go quickly from automated analysis of multiple sets of acquired topographic data to the reporting of precise results. https://www.leica-microsystems.com//science-lab/for-rapid-accurate-reporting-of-results/ Wed, 08 May 2019 22:00:00 +0000 M.S. Albert S. Laforet, PhD James DeRose https://www.leica-microsystems.com/24942 Quantitative Imaging The Phasor analysis approach to analyze fluorescence lifetime does not require any fitting. Phasor FLIM (fluorescence lifetime imaging microscopy) provides a 2D graphical view of lifetime distributions. This graphical view enables any observer to distinguish and separate different lifetime populations within a FLIM image rapidly. The interpretation of phasor FLIM distributions is straightforward. Multiple molecular species are resolved within a single pixel, because every species has a specific phasor. https://www.leica-microsystems.com//science-lab/phasor-analysis-for-flim-fluorescence-lifetime-imaging-microscopy/ Tue, 07 May 2019 22:00:00 +0000 Dr. Giulia Ossato https://www.leica-microsystems.com/24969 Surgical Microscopy Experienced ophthalmic surgeons explain how they perform cataract surgery with high efficiency using the Proveo 8 ophthalmic microscope from Leica Microsystems. The sophisticated illumination system of the Proveo 8 provides the surgeons with an excellent red reflex during the entire procedure. They describe how they get the image quality they need during hydro dissection, phacoemulsification or cataract extraction using Proveo 8. https://www.leica-microsystems.com//science-lab/surgeons-describe-how-proveo-8-helps-to-streamline-their-workflow/ Thu, 02 May 2019 22:00:00 +0000 https://www.leica-microsystems.com/24931 Coherent Raman Scattering (CRS) Leica Microsystems‘ Workshop on Coherent Raman Scattering (CRS) Microscopy attracted 30 enthusiastic participants from all over the world to beautiful Konstanz, Germany! The group enjoyed a number of great presentations, posters, and intense hands-on sessions on the latest Leica technology, including Coherent Anti-Stokes and Stimulated Raman Scattering microscopy (CARS/SRS), the FALCON lifetime imaging platform, and the THUNDER imagers. It was amazing to see a community of CARS/SRS users developing the technology in a physics lab and then using it for applications in biology, medicine, pharma, and much more. https://www.leica-microsystems.com//science-lab/summary-on-coherent-raman-scattering-crs-microscopy-workshop-in-konstanz-germany/ Sun, 28 Apr 2019 22:00:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/24929 Confocal Microscopy Cells interact with their environment. Not only on the base of chemical signals, but also by sensing and modifying mechanical properties of the extracellular matrix. The research goal of Dr. Bo Sun’s group at Oregon State University is to understand such interactions. One of their tools is visualization of matrix properties with a TCS SPE confocal microscope. https://www.leica-microsystems.com//science-lab/visualize-mechanical-interactions-of-cancer-cells/ Tue, 16 Apr 2019 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24781 Confocal Microscopy Neuroscience Structural information is key when one seeks to understand complex biological systems, and one of the most complex biological structures is the vertebrate central nervous system. To image a complete brain dissected from a developing zebrafish, one would need to cover a field of some ten square millimeters at a depth in the millimeter range. Usually, low magnification lenses do not provide sufficient resolution to reveal the intricate structural interactions in nervous tissue. Additionally, due to scattering processes, the depth at which one can image within dense biological tissue using a confocal microscope is generally restricted to approximately 10 microns. https://www.leica-microsystems.com//science-lab/zebrafish-brain-whole-organ-imaging-at-high-resolution/ Mon, 08 Apr 2019 22:00:00 +0000 Ryan E. Robinson, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24735 Laser Microdissection Lectin microarray (LMA) is a highly sensitive technology used to obtain the global glycomic profiles of endogenous glycoproteins in biological samples including formalin-fixed paraffin-embedded tissue sections. Here, we describe an effective method for cell type-selective glycomic profiling of tissue fragments collected by laser microdissection (LMD) under fluorescent histochemical visualization. We optimized each step of histochemical staining and confirmed the reliability and validity of glycomic profiling. Using the optimized procedure, glycomic profiles were obtained with 0.5 mm2 of stained thymic sections (5-μm-thick) from 8-week-old C57BL/6J male mice. https://www.leica-microsystems.com//science-lab/analysis-of-tissue-sections-assisted-by-fluorescence-laser-microdissection/ Tue, 26 Mar 2019 23:00:00 +0000 https://www.leica-microsystems.com/24749 Digital Microscopy Save time for your 2D and 3D analysis. Watch this video to learn about the new user interface, LAS X.next, for the DVM6 digital microscope. The video demonstrates how to make a fast Z-Stack with a few clicks. https://www.leica-microsystems.com//science-lab/how-to-make-a-fast-z-stack/ Mon, 25 Mar 2019 23:00:00 +0000 PhD James DeRose https://www.leica-microsystems.com/24755 Live-Cell Imaging The wound healing assay, often used as a synonym for scratch assay or migration assay, is an important tool for the evaluation and measurement of cell migration, whether single cells or cell populations. Cell migration is tightly controlled during ontogenetic processes and at times can get out of control, e.g., during cancer development and progression. https://www.leica-microsystems.com//science-lab/how-can-paula-help-streamline-the-workflow-for-my-wound-healing-assay/ Mon, 18 Mar 2019 23:00:00 +0000 Dr. Karin Schwab https://www.leica-microsystems.com/24754 Optical Coherence Tomography The aim of this study was to describe intraoperative optical coherence tomography (OCT) as an adjunctive device for peripheral synechiolysis in a post-penetrating keratoplasty patient. https://www.leica-microsystems.com//science-lab/use-of-enfocus-intraoperative-oct-to-overcome-difficulties-in-visualization-during-synechiolysis/ Sun, 17 Mar 2019 23:00:00 +0000 https://www.leica-microsystems.com/27638 Laser Microdissection Brain research: Single neuron excision for RNA analysis - Improve your workflow with laser microdissection - Parkinson’s disease is a common progressive neurodegenerative disorder. It is connected with cell death of dopamine-releasing neurons in the substantia nigra. Differences in gene expression patterns between individual dopamine-releasing neurons of disease affected and healthy individuals allow defining target genes for therapies. For RNA analysis, single cell resolution is crucial, as investigating the whole tissue is meaningless. Analyzing mixtures of dopamine-releasing neurons and all other brain cells distorts the result. https://www.leica-microsystems.com//science-lab/how-to-improve-your-rna-analysis-workflow-with-laser-microdissection/ Sun, 17 Mar 2019 10:30:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/24696 Confocal Microscopy The SP detector from Leica Microsystems denotes a compound detection unit for point scanning microscopes, in particular confocal microscopes. The SP detector splits light into up to 5 spectral bands. The bands are independent and continuously tunable within the full visible spectrum. The light in each band is detected by a light sensor: a photomultiplier tube (PMT) or a Hybrid Detector (HyD). https://www.leica-microsystems.com//science-lab/what-is-a-spectral-detector-sp-detector/ Sun, 10 Mar 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24698 Confocal Microscopy A resonant scanner is a type of galvanometric mirror scanner that allows fast image acquisition with single-point scanning microscopes (true confocal and multiphoton laser scanning). High acquisition speed is required to track fast processes, especially in living samples. It has the additional benefit of providing a better fluorescence signal and reduced photobleaching. https://www.leica-microsystems.com//science-lab/what-is-a-resonant-scanner/ Sun, 10 Mar 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/27569 Confocal Microscopy Understanding the sub-cellular mechanisms in carcinogenesis is of crucial importance for cancer treatment. Popular cellular models comprise cancer cells grown as monolayers. But this approach disregards the three-dimensional (3D) interaction of tumor cells with their surrounding microenvironment. To understand the development and progression of malignancy in a close to nature context, the characterization of cancer microenvironments is crucial. https://www.leica-microsystems.com//science-lab/improve-3d-cell-biology-workflow-with-digital-light-sheet-microscopy/ Sun, 10 Mar 2019 08:53:00 +0000 https://www.leica-microsystems.com/27595 Confocal Microscopy Visualize the ultrastructure of the glomerular filtration barrier. The combination of optical clearing and nanoscopy allows to study the glomerular filtration barrier at the nanoscale. The Leica TCS SP8 STED super-resolution microscope equipped with the STED WHITE Glycerin objective sheds light on details deep inside the specimen with 3D STED. https://www.leica-microsystems.com//science-lab/improve-workflow-in-kidney-research-with-3d-sted-deep-nanoscopy/ Sat, 09 Mar 2019 08:34:00 +0000 https://www.leica-microsystems.com/24707 EM Sample Preparation The EM ICE is the only available high-pressure freezer combining light stimulation with fast cryo fixation, a module mostly used for optogenetic approaches in neurobiology studies. However, this approach can be also very interesting for pharmaceutical or cosmetic industries to study the influence of UV light exposure on their target structures. This article shows an example on how UV light exposure changes the ultrastructure of sun screen lotion. https://www.leica-microsystems.com//science-lab/cryo-sem-analysis-of-uv-light-stimulated-sun-screen-lotion/ Wed, 06 Mar 2019 23:00:00 +0000 PhD Dietmar Pum, PhD Saskia Mimietz-Oeckler https://www.leica-microsystems.com/24709 Confocal Microscopy A field-of-view scanner is an assembly of galvanometric scanning mirrors used in single-point confocal microscopes that offer the correct optical recording of large field sizes. The field-of-view scanner employs a three-mirror concept that offers superior homogeneity in illumination compared to classical two-mirror scanners, without compromising high scanning speed. The galvanometric scanners are controllable in speed and position, allowing zoom and pan functions as well as tuning the scan frequency. They also allow resting point illumination as is required for, e.g., FCS-measurements. https://www.leica-microsystems.com//science-lab/what-is-a-field-of-view-scanner/ Wed, 06 Mar 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24711 Confocal Microscopy A HyD is a sensor for detecting light (photons). It is a compound (hybrid) of two technologies: vacuum tubes, like those used in a photomultiplier tube (PMT), and semiconductor microelectronics, like an avalanche photodiode (APD). The Hybrid Detector combines essentially the advantages of both approaches. It is fast, shows very little noise, and offers a high dynamic range. For a comparison see references 1, 2, 3, and 4. The HyD is the best suited sensor for most confocal and multiphoton applications. https://www.leica-microsystems.com//science-lab/what-is-a-hybrid-detector-hyd/ Wed, 06 Mar 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24713 Confocal Microscopy The field number (FN) for optical microscopes indicates the field of view (FOV). It corresponds to the area in the intermediate image that is observable through the eyepieces. Although, we cannot observe very large fields at once, the human eye can scan and integrate structural features over the full field. Furthermore, it is essential, that field size and resolution fit to the capabilities of the human eye. https://www.leica-microsystems.com//science-lab/resolved-field-number-rfn/ Wed, 06 Mar 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24534 Widefield Microscopy The present study describes the optimal growth conditions for the proliferation and differentiation of bovine bronchial epithelial cells (BBECs). The cells are grown at an air-liquid interface (ALI) in a serum-free medium to produce an airway epithelial cell (AEC) model. The AEC model accurately mimics the in vivo bovine respiratory epithelium. https://www.leica-microsystems.com//science-lab/how-to-develop-an-airway-epithelial-cell-model/ Wed, 06 Mar 2019 23:00:00 +0000 Eugenia Petoukhov https://www.leica-microsystems.com/24637 Widefield Microscopy Fluorescence Microscopy Live-Cell Imaging Stereo Microscopy So far, widefield microscopy was not suitable to image larger volumes, since the contrast of the recorded fluorescence image is reduced by the background (BG) mainly originating from out of focus regions of the recorded volume. BG reduces the effective dynamic range, the maximal possible signal to noise ratio (SNR), of the imaging system. https://www.leica-microsystems.com//science-lab/thunder-technology-note/ Tue, 05 Mar 2019 23:00:00 +0000 Dr. Jan Schumacher, Louise Bertrand https://www.leica-microsystems.com/24700 Widefield Microscopy Stereo Microscopy Fluorescence Microscopy Live-Cell Imaging https://www.leica-microsystems.com//science-lab/thunder-technology-note/thank-you-for-your-registration/ Tue, 05 Mar 2019 23:00:00 +0000 https://www.leica-microsystems.com/24702 EM Sample Preparation Analyzing purified protein structures in a cellular context is key to understanding how proteins and protein complexes function. Thermo Fisher Scientific and Leica Microsystems developed the integrated cryo-tomography workflow to enable high resolution structural analysis, done in a controlled and effective way, of proteins in their native environments. In this webinar, the speakers will explain the unique features of the hardware and software integration and guide you step-by-step through the workflow. https://www.leica-microsystems.com//science-lab/changing-the-way-we-look-at-the-inner-workings-of-cells/ Sun, 03 Mar 2019 23:00:00 +0000 https://www.leica-microsystems.com/24715 EM Sample Preparation Optimal structural preservation of tissue can only be obtained by high pressure freezing. However, preparing the samples in optimal conditions is challenging. This article explains in detail how to dissect, high pressure freeze and freeze substitute Droshophila tissue sample to obtain high structural integrity for subsequent electron microscopic analysis. https://www.leica-microsystems.com//science-lab/superior-ultrastructural-preservation-and-structural-contrast-in-drosophila-tissue/ Sun, 03 Mar 2019 23:00:00 +0000 PhD Zulfeqhar Syed, PhD Christopher Bleck https://www.leica-microsystems.com/27643 Laser Microdissection Brain Research: Collect pure starting material for proteomics - Improve your workflow with Laser Microdissection - Many brain diseases result from protein malfunction, misfolding and agglutination. For this reason protein analysis is the key to understanding causes of, and discovering therapies for, many cerebral defects. https://www.leica-microsystems.com//science-lab/how-to-improve-your-alzheimer-protein-analysis-with-laser-microdissection/ Sat, 02 Mar 2019 10:53:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/24694 Confocal Microscopy A Tandem Scanner is an assembly of two different types of scanning together in one system for true confocal point scanning. The Tandem Scanner consists of a three-mirror scanning base with the x-scanner exchangeable with a motorized device. This combination allows scanning of large areas with high scan-resolution by a FOV-scanner and of very fast processes by a resonant scanner, both within the same instrument. https://www.leica-microsystems.com//science-lab/what-is-a-tandem-scanner/ Thu, 28 Feb 2019 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/24692 Quantitative Imaging An international consortium of researchers has found a new mutation in a gene harboring a reticulum-to-cytoplasm zinc transporter. The gene is called ZIP7 and it is responsible for the early onset of infections in patients. These findings led them to uncover the role of ZIP7 in B cell development. To elucidate the cellular function of ZIP7, researchers used CRISPR-Cas9 mutagenesis to recreate the observed mutations in mice. They then studied primary cells with functional imaging and zinc biosensors to corroborate the mutations and see how cells handle zinc. https://www.leica-microsystems.com//science-lab/researchers-use-functional-imaging-to-elucidate-the-role-of-cellular-zn2-in-human-immunity/ Wed, 27 Feb 2019 23:00:00 +0000 Dr. Luis Alvarez https://www.leica-microsystems.com/24680 Live-Cell Imaging Many cell-based experiments require cells in a certain state. This can include their morphology,  fluorescent protein expression, and the confluency of the cell layer. Researchers check their cells regularly to determine these attributes. Confluency estimation can be very difficult! This guess work is biased by every individual researcher, and can result in lab error.  https://www.leica-microsystems.com//science-lab/confluency-check-with-paula-cell-imager/ Wed, 20 Feb 2019 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/24669 Super-Resolution When it comes to STED Nanoscopy, keeping high signal-to-noise is key to achieve the best possible resolution in fixed and living cells. This can be challenging in the case of experiments in 3D and/or with time series, where the sample undergoes many rounds of image acquisition and photobleaching becomes an issue. If fluorophores were completely immune to photobleaching, it should be possible to perform STED indefinitely using the same molecules over and over. In practice, one performs STED with the best available fluorophores in terms of brightness and photostability (Grimm, Muthusamy et al. 2017), and at high labeling densities. However, there is a clever alternative to come closer to the ideal situation: if “fresh” fluorophores replenish the sample in each round of STED, imaging will take place with intact fluorophores every time. https://www.leica-microsystems.com//science-lab/extending-nanoscopy-possibilities-with-sted-and-exchangeable-fluorophores/ Tue, 12 Feb 2019 23:00:00 +0000 Dr. Julia Roberti https://www.leica-microsystems.com/24656 Light Sheet Microscopy Sample handling is an important topic in the context of Light Sheet Microscopy. The TCS SP8 DLS integrates Light Sheet technology into an inverted confocal platform and can hence make use of general principles concerning sample mounting and xy-stage functionality. We describe a set of mounting frames that allow preparation of a higher number of samples, also in the context of potentially harmful mounting media like BABB. https://www.leica-microsystems.com//science-lab/using-mounting-frames-for-dls-sample-preparation/ Sun, 10 Feb 2019 23:00:00 +0000 Simone Diehl, Dr. Petra Haas https://www.leica-microsystems.com/27591 Laser Microdissection DNA mutations lead to abnormal proteins or missing functional proteins, which can cause cells to multiply uncontrollably and become cancerous. To find and understand the underlying mutation for a specific cancer type, extraction of pure tumor material is extremely important. This is challenging especially if only a limited amount of cancerous tissue is available (small metastasis, for example). Laser Microdissection allows for the extraction of purely cancerous tissue without contaminating your sample with healthy cells. DNA sequencing will lead to valid and reproducible results helping to identify the cancer causing DNA mutation easily as no masking by contaminating healthy cells can occur. https://www.leica-microsystems.com//science-lab/how-to-improve-your-dna-mutation-analysis-workflow-with-laser-microdissection/ Fri, 08 Feb 2019 12:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/20488 Light Sheet Microscopy The TCS SP8 DLS from Leica Microsystems is an innovative concept to integrate the Light Sheet Microscopy technology into the confocal microscope. Due to its unique optical architecture samples can be mounted on standard glass bottom petri dishes and require only little adaptations when compared to conventional mounting procedures, as for confocal imaging. https://www.leica-microsystems.com//science-lab/using-a-rotation-device-for-dls-sample-mounting/ Tue, 22 Jan 2019 23:00:00 +0000 Dr. Petra Haas, Elena Remacha Motta, Laia Ortiz Lopez, Dr. Bruno Cossermelli Vellutini, Dr. Pavel Tomancak, Dr. Emmanuel Reynaud https://www.leica-microsystems.com/24585 Confocal Microscopy Blood vessels transport vital nutrients and oxygen to all the cells in the body. Guided by a complex signaling network, endothelial cells sprout, proliferate, and migrate to form those vessels. One of the processes of vascular formation is angiogenesis, the growth of new vessels from pre-existing ones. Our current knowledge of the mechanisms controlling angiogenesis and vascular stability is still limited. However, these mechanisms play important roles in the pathophysiology of cancer, diabetes and stroke. Understanding the factors that control healthy and unhealthy blood vessel development is fundamental to prevent and fight such diseases. https://www.leica-microsystems.com//science-lab/a-molecular-link-between-cell-migration-and-vascular-disease/ Wed, 19 Dec 2018 23:00:00 +0000 Dr. Julia Roberti https://www.leica-microsystems.com/24579 EM Sample Preparation Aluminum (Al) alloys play an important role in the production of aircraft and vehicles, as well as products in other industries. Defects present in the Al alloy used for the production of aircraft, vehicles, or other products can have a significant effect on their quality, performance, and lifetime. Characterization of Al alloys in 3D can lead to a better understanding of defect formation and how to minimize or eliminate it. https://www.leica-microsystems.com//science-lab/quick-3d-imaging-of-alloy-defects/ Thu, 13 Dec 2018 23:00:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/24562 Confocal Microscopy The Cell Atlas, a part of the Human Protein Atlas, was created by the group of Prof. Emma Lundberg at the SciLifeLab, KTH Royal Institute of Technology, in Stockholm, Sweden. Currently, she is a visiting professor at Stanford University through the support of the Chan Zuckerberg Initiative. The Cell Atlas was created, in large part, using data acquired with Leica confocal instruments. In the scope of the Kaggle competition regarding the Human Protein Atlas Image Classification Prof. Lundberg gave an interview to Dr. Constantin Kappel from Leica Microsystems. https://www.leica-microsystems.com//science-lab/kaggle-competition-for-multi-label-classification-of-cell-organelles-in-proteome-scale-human-protein-atlas-data/ Sun, 09 Dec 2018 23:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/24564 EM Sample Preparation Array tomography (AT) is a 3D image reconstruction technique for high resolution, quantitative analysis of biological structures. For optimal results, ultrathin and ordered sections are an absolute requirement. Watch this webinar and get tips and tricks to optimize the workflow of your array tomography. https://www.leica-microsystems.com//science-lab/array-tomography-for-sem-3d-reconstruction/ Thu, 06 Dec 2018 23:00:00 +0000 Robert Ranner https://www.leica-microsystems.com/24565 EM Sample Preparation Capturing the intricate changes in fine structure or in cell dynamics with conventional cryo solutions can be challenging sometimes. Leica Microsystems has developed a new cryo platform, the Leica EM ICE, to help expand the current limits of cryofixation. Watch this free webinar and learn how the Leica EM ICE combines speed, reliability and flexibility to facilitate research in various scientific fields. https://www.leica-microsystems.com//science-lab/expanding-the-limits-of-electron-microscopy-sample-preparation/ Thu, 06 Dec 2018 23:00:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/24560 Super-Resolution Despite the emergence of new imaging methods in recent years, true 3D resolution is still achieved by Confocal Laser Scanning Microscopy (CLSM). Through a combination of novel, extremely fast scanning methods with high sensitivity, low noise detectors and simultaneous multi-spectral data acquisition, Leica’s confocal imaging has now been extended to the point that previously inaccessible dynamic and spectral ranges became accessible. https://www.leica-microsystems.com//science-lab/see-more-than-just-your-image/ Tue, 04 Dec 2018 23:00:00 +0000 https://www.leica-microsystems.com/24557 Super-Resolution The Endoplasmic Reticulum (ER) is a dynamic, reticular or net-like structure consisting of tubules and cisternae that extend throughout the cell and occupy a large portion of the cytoplasm. In recent years, it has become clear that the ER is not a static organelle. Instead the ER is dynamic, forming contacts with nearly every intracellular membrane. Recent innovations that improve image resolution in light microscopy have advanced our ability to study the dynamics of the endoplasmic reticulum and the formation of ER contact sites with other organelles. https://www.leica-microsystems.com//science-lab/dynamic-endoplasmic-reticulum/ Fri, 30 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/24510 Confocal Microscopy Yutaka Kodama is Associate Professor at the Biosciences Education Research Center, Utsunomiya University, Japan. He graduated from Saga University, Faculty of Agriculture, in 2004 and received his PhD from the Graduate School of Biological Science at the Nara Institute of Science and Technology in 2007. https://www.leica-microsystems.com//science-lab/you-shall-not-pass-time-gated-detection-takes-care-of-endogenous-fluorescence-in-plant-research/ Tue, 13 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/24508 Super-Resolution Confocal Microscopy Live-Cell Imaging Reliable interpretation and quantification of cellular features in fluorescence microscopy requires an accurate estimate of microscope resolution. This is typically obtained by measuring the image of a nonbiological proxy for a point-like object, such as a fluorescent bead. Although appropriate for confocal microscopy, bead-based measurements are problematic for stimulated emission depletion microscopy and similar techniques where the resolution depends critically on the choice of fluorophore and acquisition parameters. In this article, we demonstrate that for a known geometry (e.g., tubules), the resolution can be measured in situ by fitting a model that accounts for both the point spread function (PSF) and the fluorophore distribution. https://www.leica-microsystems.com//science-lab/simultaneously-measuring-image-features-and-resolution-in-live-cell-sted-images/ Mon, 12 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/20689 Multiphoton Microscopy Today’s life science research focusses on complex biological processes, such as the causes of cancer and other human diseases. A deep look into tissues and living specimens is vital to understanding the conditions and mechanisms in cells and finding answers to crucial questions challenging the life sciences. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-tunable-multicolor-deep-in-vivo-imaging/ Thu, 01 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/20671 Confocal Microscopy Programmed cell death (PCD)-triggered degradation of plant tapetum is essential for microspore development and pollen coat formation; however, little is known about the cellular mechanism regulating tapetal PCD. Here, we demonstrate that Rab7-mediated vacuolar transport of tapetum degradation-related cysteine proteases is crucial for tapetal PCD and pollen development in Arabidopsis (Arabidopsis thaliana), with the following evidence: (1) The monensin sensitivity1 (mon1) mutants, which are defective in Rab7 activation, showed impaired male fertility due to a combined defect in both tapetum and male gametophyte development. https://www.leica-microsystems.com//science-lab/monensin-sensitivity1-mon1calcium-caffeine-zinc-sensitivity1-ccz1/ Mon, 22 Oct 2018 22:00:00 +0000 https://www.leica-microsystems.com/20653 Laser Microdissection Laser microdissection is a tool for the isolation of homogenous cell populations from their native niches in tissues to downstream molecular assays. Beside its routine use for fixed tissue sections, laser microdissection may be applied for live cell isolation. Unlike other well-established and widely used techniques for live cell isolation and single cell cloning—such as FACS, MACS, cloning by limited dilution, and so on—laser microdissection allows for capturing live cells and cell colonies without their detachment from the carrier. https://www.leica-microsystems.com//science-lab/live-cell-isolation-by-laser-microdissection/ Tue, 16 Oct 2018 22:00:00 +0000 https://www.leica-microsystems.com/20642 EM Sample Preparation The optimization of high resolution, 3-dimensional (3D), sub-cellular structure analysis with array tomography using an automated serial sectioning solution, achieving a high section density on the carrier substrate, is described in this report. https://www.leica-microsystems.com//science-lab/high-resolution-array-tomography-with-automated-serial-sectioning/ Sun, 14 Oct 2018 22:00:00 +0000 Robert Ranner, PhD James DeRose https://www.leica-microsystems.com/20511 Super-Resolution Confocal Laser Scanning Microscopy (CLSM) is the standard for true 3D resolved fluorescence imaging. Fast optical sectioning using flexible scanning strategies in combination with simultaneous multi-colour, high sensitivity and low noise signal detection provides maximum resolution in the spatial and temporal domain. In combination with modern approaches to image information extraction this helps the researcher to mine as much information as possible from the images acquired. Image information extraction refers to intelligent procedures for image enhancement using a priori knowledge from the imaging system. From simple glare control and optical development to intelligent and ingenious model extraction, there are many ways to see more than just the image. https://www.leica-microsystems.com//science-lab/how-to-extract-image-information-by-adaptive-deconvolution/ Tue, 18 Sep 2018 22:00:00 +0000 Dr. Jürgen Reymann https://www.leica-microsystems.com/20495 Laser Microdissection Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches require samples comprising a minimum of thousands of cells to provide in-depth profiling. Here, we report the development of a nanoPOTS (nanodroplet processing in one pot for trace samples) platform for small cell population proteomics analysis. NanoPOTS enhances the efficiency and recovery of sample processing by downscaling processing volumes to <200 nL to minimize surface losses. https://www.leica-microsystems.com//science-lab/nanodroplet-processing-platform-for-deep-and-quantitative-proteome-profiling-of-10-100-mammalian-cells/ Wed, 12 Sep 2018 22:00:00 +0000 https://www.leica-microsystems.com/20352 EM Sample Preparation Physical porosity in rocks, like shale and carbonate, has a large effect on the their storage capacity. The pore geometries also affect their permeability. Imaging the visible pore space provides insights into the physical pore space, pore geometries, and the associated mineral and organic matter phases relevant for storage and transport. For example, organic-matter porosity is an important property of mature organic-rich shales providing storage capacity for liquids and gases, like hydrocarbons. Imaging the microstructure is the only technique which delivers data giving direct insight into the organic matter porosity. However, imaging porosity in fine-grained shale as well as carbonate rocks in a representative way at high resolution is challenging. https://www.leica-microsystems.com//science-lab/macroscale-to-nanoscale-pore-analysis-of-shale-and-carbonate-rocks/ Tue, 04 Sep 2018 22:00:00 +0000 PhD Jop Klaver, PhD Joyce Schmatz, MSc Mingze Jiang, PhD James DeRose https://www.leica-microsystems.com/20475 Super-Resolution Malaria is a life-threatening disease transmitted through the bites of mosquitoes infected with protozoan parasites. The most common and dangerous type of malaria is caused by the parasite Plasmodium falciparum. Malaria raises serious concerns because half of the world’s population is at risk and there are no effective vaccines available to date. WHO reported 216 million cases in 2016 [1], a half-million deaths, and an economic burden exceeding the billion dollar range. https://www.leica-microsystems.com//science-lab/observing-malaria-infection-at-the-right-spot-in-the-human-host/ Wed, 29 Aug 2018 22:00:00 +0000 Dr. Julia Roberti https://www.leica-microsystems.com/20418 Coherent Raman Scattering (CRS) Multiphoton Microscopy The artery wall is equipped with a water permeation barrier that allows blood to flow at high pressure without significant water leak. The precise location of this barrier is unknown despite its importance in vascular function and its contribution to many vascular complications when it is compromised. Herein we map the water permeability in intact arteries, using coherent anti-Stokes Raman scattering (CARS) microscopy and isotopic perfusion experiments. Generation of the CARS signal is optimized for water imaging with broadband excitation. https://www.leica-microsystems.com//science-lab/direct-visualization-of-the-arterial-wall-water-permeability-barrier-using-cars-microscopy/ Wed, 22 Aug 2018 22:00:00 +0000 https://www.leica-microsystems.com/20359 Quantitative Imaging Förster (Fluorescence) Resonance Energy Transfer (FRET) has become a powerful tool to study protein-protein interactions and signal transduction in living cells. FRET is commonly read out either by detecting the ratio of the donor and acceptor intensities (sensitized emission) or by detecting the excited state lifetime of the donor, which decreases with increasing FRET (Fluorescence Lifetime IMaging or FLIM). https://www.leica-microsystems.com//science-lab/precise-and-lightning-fast-spectral-fluorescence-lifetime-imaging-at-video-rate-integrated-in-a-high-end-confocal-microscope/ Tue, 07 Aug 2018 22:00:00 +0000 PhD Kees Jalink https://www.leica-microsystems.com/20357 Super-Resolution Molecular interactions are key in cellular signalling. They are often ruled or rendered by the mobility of the involved molecules. https://www.leica-microsystems.com//science-lab/super-resolved-sted-spectroscopy/ Mon, 06 Aug 2018 22:00:00 +0000 Prof. Christian Eggeling https://www.leica-microsystems.com/20350 Widefield Microscopy Bloodsucking insects play a major role in the transmission of pathogens that cause major tropical diseases. Their capacity to transmit these diseases is directly associated with the availability and turnover of energy sources. Proline is the main readily-mobilizable fuel of the tsetse fly, which is the vector of sub-species of Trypanosoma brucei parasites that cause human sleeping sickness and are partly responsible for animal trypanosomiasis (Nagana disease) in sub-Saharan Africa. https://www.leica-microsystems.com//science-lab/proline-metabolism-is-essential-for-trypanosoma-brucei-brucei-survival-in-the-tsetse-vector/ Sun, 29 Jul 2018 22:00:00 +0000 https://www.leica-microsystems.com/20323 Confocal Microscopy The Hybrid Photodetectors (HyD) are! Why that is the case is explained in this short Science Lab article. https://www.leica-microsystems.com//science-lab/which-sensor-is-the-best-for-confocal-imaging/ Mon, 16 Jul 2018 22:00:00 +0000 Dr. Rolf T. Borlinghaus, Holger Birk https://www.leica-microsystems.com/20306 Super-Resolution Alison Dun is the postdoctoral facility manager for the Edinburgh Super-Resolution Imaging Consortium (ESRIC), Heriot-Watt University, Edinburgh, UK. She has used a large range of microscope techniques during her PhD work. In a video published by BBC News, Alison Dun explains how STED (stimulated emission depletion) microscopes help researchers in the fight against cancer by getting a better understanding of the processes that go wrong in the human body, thus giving hope for future treatments. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-at-the-forefront-of-cancer-research/ Thu, 05 Jul 2018 22:00:00 +0000 PhD Alison Dun, Dr. Julia Roberti https://www.leica-microsystems.com/20297 Confocal Microscopy SP8 FALCON (FAst Lifetime CONtrast) is the fast, and fully integrated platform for fluorescence lifetime imaging (FLIM) and analysis. It enables the observation of fast dynamic cellular events with lifetime contrasts between fluorescent molecules down to sub-second time scales. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-flim-for-high-speed-investigation-of-molecular-interactions/ Wed, 04 Jul 2018 22:00:00 +0000 Dr. Johanna Berndt https://www.leica-microsystems.com/15127 Basics in Microscopy Digital Microscopy Stereo Microscopy One important criterion concerning the performance of an optical microscope is magnification. This report will offer digital microscopy users helpful guidelines to determine the useful range of magnification values. https://www.leica-microsystems.com//science-lab/what-does-300001-magnification-really-mean/ Mon, 25 Jun 2018 22:00:00 +0000 PhD James DeRose, MSc Michael Doppler https://www.leica-microsystems.com/19763 Metallography Quality Assurance Forensics The advantages of a 2-in-1 materials analysis solution combining optical microscopy and laser induced breakdown spectroscopy (LIBS) for simultaneous visual and chemical inspection are described in this report. The basic principles of the 2-in-1 solution and a comparison between it and other common materials analysis methods, such scanning electron microscopy (SEM), are explained to demonstrate how a rapid, efficient workflow is achieved. A 2-in-1 analysis solution can reduce significantly the cost and time for obtaining material image and composition data. Such data are instrumental in assuring quality and reliability to make confident decisions quickly during production, quality control, failure analysis, and research and development in industries and fields, such as automotive and metallurgy. https://www.leica-microsystems.com//science-lab/see-the-structure-with-microscopy-know-the-composition-with-laser-spectroscopy/ Mon, 11 Jun 2018 13:39:00 +0000 PhD James DeRose, Dr. Kay Scheffler https://www.leica-microsystems.com/20208 Quantitative Imaging Fluorescence Lifetime Imaging (FLIM) in combination with Förster Resonance Energy Transfer (FRET) has proven to be very beneficial for investigations in biomedical research for a wide range of structural elements and dynamic changes in cells. FRET allows to monitor molecular interactions, as the FRET signal depends strongly on the distance of the two FRET partners. This allows to investigate interaction of molecules, like ligand-receptor pairs, protein-protein interactions or interactions of effectors with DNA. https://www.leica-microsystems.com//science-lab/flim-fret-and-biosensors-versatile-tools-for-biomedical-research/ Sun, 10 Jun 2018 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/20202 Super-Resolution Neurons react to learning and memory by activating synaptic connections. The mechanisms behind this fundamental process are complex and poorly understood. Researchers at Thomas Jefferson University have found that neuron plasticity operates in a “digital” fashion through nanomodules of discrete size that multiply and strengthen neuronal connections upon stimulation. This breakthrough was published on April 23rd in the journal Nature Neuroscience. https://www.leica-microsystems.com//science-lab/researchers-find-a-digital-mechanism-behind-neuronal-changes-from-learning/ Mon, 04 Jun 2018 22:00:00 +0000 Dr. Martin Hruska, Dr. Julia Roberti https://www.leica-microsystems.com/20180 Coherent Raman Scattering (CRS) Confocal Microscopy The chemical basis for the alteration of the refractive properties of an intraocular lens with a femtosecond laser was investigated. Three different microscope setups have been used for the study: Laser Induced Fluorescence (LIF) microscopy, Raman microscopy and coherent anti-Stokes Raman Scattering (CARS) microscopy. https://www.leica-microsystems.com//science-lab/chemical-basis-for-alteration-of-an-intraocular-lens-using-a-femtosecond-laser/ Sun, 27 May 2018 22:00:00 +0000 https://www.leica-microsystems.com/20175 Confocal Microscopy Widefield Microscopy The myelination of axons is essential for neuronal wiring and normal nervous system functions. In the peripheral nervous system, Schwann cells (SCs) form myelin sheaths around axons during nerve development. Such myelination is compromised in a number of diseases. Hence, identification and understanding of the key pathways regulating SC development and myelinogenesis are essential for therapeutic progress. Here we uncover two separate roles of the cellular signaling node mTORC1 (mechanistic target of rapamycin complex 1) for regulating the development of SCs and subsequently the growth of myelin sheaths. Moreover, we demonstrate that defective SCs possess a remarkable plasticity to remyelinate axons via mTORC1. Thus, manipulating mTORC1 activity in diseased SCs could be therapeutically beneficial. https://www.leica-microsystems.com//science-lab/mtorc1-promotes-proliferation-of-immature-schwann-cells-and-myelin-growth-of-differentiated-schwann-cells/ Wed, 16 May 2018 22:00:00 +0000 https://www.leica-microsystems.com/20149 Confocal Microscopy Development of stomata, valves on the plant epidermis for optimal gas exchange and water control, is fine-tuned by multiple signaling peptides with unique, overlapping, or antagonistic activities. EPIDERMAL PATTERNING FACTOR1 (EPF1) is a founding member of the secreted peptide ligands enforcing stomatal patterning. Yet, its exact role remains unclear. Here, we report that EPF1 and its primary receptor ERECTA-LIKE1 (ERL1) target MUTE, a transcription factor specifying the proliferation-to-differentiation switch within the stomatal cell lineages. https://www.leica-microsystems.com//science-lab/autocrine-regulation-of-stomatal-differentiation-potential-by-epf1-and-erecta-like1-ligand-receptor-signaling/ Sun, 13 May 2018 22:00:00 +0000 https://www.leica-microsystems.com/20142 Multiphoton Microscopy Confocal Microscopy Zebrafish testis has become a powerful model for reproductive biology of teleostean fishes and other vertebrates and encompasses multiple applications in applied and basic research. Many studies have focused on 2D images, which is time consuming and implies extrapolation of results. Three-dimensional imaging of whole organs recently became an important challenge to better understand their architecture and allow cell enumeration. https://www.leica-microsystems.com//science-lab/high-resolution-3d-imaging-of-whole-organ-after-clearing/ Tue, 08 May 2018 22:00:00 +0000 https://www.leica-microsystems.com/20127 EM Sample Preparation Precise low angle rotary shadowing with heavy metals (like platinum) can be used in transmission electron microscopy (TEM) to observe molecular details of objects previously absorbed on a thin, low grain and electron-transparent carbon film. To achieve the highest contrast and better image quality, it is essential that the coating is directional, and it is given at a precise angle toward the sample. The fine grain of the metal layers and the homogeneous thickness of the coating material all over the sample surface are also crucial requirements to achieve high quality TEM images. This requires the method of e-beam evaporation a stream of evaporated material which is very directional, extremely homogeneous, cool and fine grained. https://www.leica-microsystems.com//science-lab/visualization-of-dna-molecules/ Wed, 02 May 2018 22:00:00 +0000 Michele Giannattasio, Gisela Höflinger https://www.leica-microsystems.com/20117 Metallography When analyzing material for cleanliness testing, users normally just want a simple way to know whether the particle under scrutiny is normal debris or something more risky. Users can now “LIBS” it to acquire rapidly the composition and then move onto the next manufacturing step. https://www.leica-microsystems.com//science-lab/leica-is-mad-about-libs/ Sun, 22 Apr 2018 22:00:00 +0000 Mike Tjepkema, PhD James DeRose https://www.leica-microsystems.com/19991 Basics in Microscopy Digital Microscopy Microscopy has become easier than ever before. Leica Microsystems offers state-of-the-art microscopes that allow to start microscopy work without a lot of training and expert skills. https://www.leica-microsystems.com//science-lab/basics-of-microscope-optics/ Tue, 10 Apr 2018 22:00:00 +0000 MSc Michael Doppler https://www.leica-microsystems.com/19998 Basics in Microscopy Microscopy has become easier than ever before. Leica Microsystems offers state-of-the-art microscopes that allow to start microscopy work without a lot of training and expert skills. Still a basic knowledge of fundamentals of optics, and the functionality of a microscope is essential for good results in microscopic work. https://www.leica-microsystems.com//science-lab/basics-of-microscope-optics/watch-the-webinar-basics-of-microscope-optics/ Tue, 10 Apr 2018 22:00:00 +0000 MSc Michael Doppler https://www.leica-microsystems.com/20109 Quantitative Imaging „Way too complicated!“ - the notorious feedback when it comes to fluorescence lifetime measurements. This will change now! New technologies and new concepts for data evaluation, all implemented in the new Leica SP8 FALCON, render fluorescence lifetime imaging (FLIM) as fuss-free as ordinary confocal imaging. And by the way: with Leica FALCON you can record frames 10 times faster compared to the classical standard. And three (or more) dimensional image stacks or time series are generated in a snap. Four channels simultaneously? No problem! And of course there are tunable excitation wavelength both visible with white light lasers (WLL) and infrared (the latter for multiphoton microscopy). That should be reason enough to delve into fluorescence lifetime imaging. The picture shows a lifetime image of a mouse embryo. Recorded in 722 stitched tiles and fitted for four separate characteristic times. Recording time ca 1 hour – compared to ca 1 day with the classical approach. https://www.leica-microsystems.com//science-lab/lifetime-a-proper-alternative/ Wed, 28 Mar 2018 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/20133 Quantitative Imaging FCS is a fluorescence-based measurement method. Fluorescent molecules passing through a strongly focused, fixed laser beam are excited for fluorescence emission. After passing a confocal pinhole, the emitted photons are registered using very sensitive detectors. https://www.leica-microsystems.com//science-lab/fcs-fluorescence-correlation-spectroscopy/ Wed, 28 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20141 Quantitative Imaging A typical application of FLIM is FLIM-FRET. FRET is a well-established technique to study molecular interactions. It scrutinizes protein binding and estimates intermolecular distances on an Angström scale as well. The SP8 FALCON system together with the integrated FRET analyzer provides FRET-efficiency and binding maps. https://www.leica-microsystems.com//science-lab/flim-fret-fluorescence-resonance-energy-transfer/ Wed, 28 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20131 Quantitative Imaging Essentially, FCS can be performed with a continuous-wave laser. Using pulsed lasers allows even more sophisticated analysis possibilities, such as time-gated FCS or Fluorescence Lifetime Correlation Spectroscopy (FLCS). Both methods make use of the additional information obtained by the simultaneous measurement of the fluorescence lifetime. https://www.leica-microsystems.com//science-lab/flcs-fluorescence-lifetime-correlation-spectroscopy/ Mon, 26 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20129 Quantitative Imaging The fluorescence lifetime is a measure of how long a fluorophore remains on average in its excited state before returning to the ground state by emitting a fluorescence photon. https://www.leica-microsystems.com//science-lab/what-is-flim-fluorescence-lifetime-imaging/ Sun, 25 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20130 Quantitative Imaging The SP8 FALCON is the ideal tool for spectral FLIM detection. No emission filters in front of the FLIM detectors are necessary. This grants a much higher flexibility to the experimental design. https://www.leica-microsystems.com//science-lab/sp-flim-spectral-fluorescence-lifetime-imaging/ Sun, 25 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20132 Quantitative Imaging FCCS (Fluorescence Cross-Correlation Spectroscopy) can be measured using the Leica TCS SP8 FCS system. Similar to FCS , it analyzes fluorescence intensity fluctuations derived from a small observation volume. https://www.leica-microsystems.com//science-lab/fccs-fluorescence-cross-correlation-spectroscopy/ Sun, 25 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20061 Live-Cell Imaging We expand upon our recent, fundamental report on solvent immersion imprint lithography (SIIL) and describe a semi-automated and high-performance procedure for prototyping polymer microfluidics and optofluidics. The SIIL procedure minimizes manual intervention through a cost-effective (∼$200) and easy-to-assemble apparatus. We analyze the procedure's performance specifically for Poly (methyl methacrylate) microsystems and report repeatable polymer imprinting, bonding, and 3D functionalization in less than 5 min, down to 8 μm resolutions and 1:1 aspect ratios. In comparison to commercial approaches, the modified SIIL procedure enables substantial cost reductions, a 100-fold reduction in imprinting force requirements, as well as a more than 10-fold increase in bonding strength. https://www.leica-microsystems.com//science-lab/solvent-immersion-imprint-lithography/ Wed, 21 Mar 2018 23:00:00 +0000 https://www.leica-microsystems.com/20037 CRISPR-Cas9 Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into induced pluripotent stem cells (iPSCs). These studies indicated that the development of biological rhythms in ESCs might be closely associated with the maintenance and differentiation of ESCs. https://www.leica-microsystems.com//science-lab/role-of-circadian-gene-clock-during-differentiation-of-mouse-pluripotent-stem-cells/ Wed, 14 Mar 2018 23:00:00 +0000 https://www.leica-microsystems.com/20021 Confocal Microscopy Pathologic electrical remodeling and attenuated cardiac contractility are featured characteristics of heart failure. Coinciding with these remodeling events is a loss of the K+ channel interacting protein, KChIP2. While, KChIP2 enhances the expression and stability of the Kv4 family of potassium channels, leading to a more pronounced transient outward K+ current, Ito,f, the guinea pig myocardium is unique in that Kv4 expression is absent, while KChIP2 expression is preserved, suggesting alternative consequences to KChIP2 loss. Therefore, KChIP2 was acutely silenced in isolated guinea pig myocytes, which led to significant reductions in the Ca²⁺ transient amplitude and prolongation of the transient duration. https://www.leica-microsystems.com//science-lab/kchip2-regulates-the-cardiac-ca2-transient-and-myocyte-contractility-by-targeting-ryanodine-receptor-activity/ Wed, 07 Mar 2018 23:00:00 +0000 https://www.leica-microsystems.com/19973 Widefield Microscopy Alpha-synuclein (α-Syn) aggregation into oligomers and fibrils is associated with dopaminergic neuron loss occurring in Parkinson’s disease (PD) pathogenesis. Compounds that modulate α-Syn aggregation and interact with preformed fibrils/oligomers and convert them to less toxic species could have promising applications in the drug development efforts against PD. Curcumin is one of the Asian food ingredient which showed promising role as therapeutic agent against many neurological disorders including PD. https://www.leica-microsystems.com//science-lab/effect-of-curcumin-analogs-ona-synuclein-aggregation-and-cytotoxicity/ Sun, 04 Mar 2018 23:00:00 +0000 https://www.leica-microsystems.com/19990 Confocal Microscopy Security is a primary concern of Leica Microsystems and Leica Microsystems’ customers that employ remote services. Leica Microsystems requires a proven remote service solution that protects against viruses and hackers and supports our intelligent instruments without major end-user modifications, while working within our current network security model and achieving official certification by a third-party security company. https://www.leica-microsystems.com//science-lab/good-to-know/it-security-whitepaper/ Wed, 28 Feb 2018 23:00:00 +0000 https://www.leica-microsystems.com/19951 Widefield Microscopy Confocal Microscopy Activation of the platelet‐derived growth factor receptor‐α (PDGFR α) signaling pathway is critically important during lung alveogenesis, the process in lung development during which alveoli are formed from the terminal alveolar sacs. Several studies have aimed to characterize the expression patterns of PDGFR α and its two ligands (PDGF‐A and ‐C) in the lung, but published analyses have been limited to embryonic and/or perinatal time points, and no attempts have been made to characterize both receptor and ligand expression simultaneously. In this study, we present a detailed map of the expression patterns of PDGFR α, PDGF‐A and PDGF‐C during the entire period of lung development, that is, from early embryogenesis until adulthood. https://www.leica-microsystems.com//science-lab/expression-analysis-of-platelet-derived-growth-factor-receptor-alpha-and-its-ligands-in-the-developing-mouse-lung/ Tue, 20 Feb 2018 23:00:00 +0000 https://www.leica-microsystems.com/19958 Super-Resolution The development of a ciliary axoneme requires the correct docking of the basal body at cytoplasmic vesicles or plasma membrane. In the multiciliated cell Paramecium, three conserved proteins, FOR20, Centrin 2, and Centrin 3 participate in this process, FOR20 and Centrin 2 being involved in the assembly of the transition zone. We investigated the function of two other evolutionary conserved proteins, OFD1 and VFL3, likely involved in this process. https://www.leica-microsystems.com//science-lab/basal-body-positioning-and-anchoring-in-the-multiciliated-cell-paramecium-tetraurelia-roles-of-ofd1-and-vfl3/ Wed, 14 Feb 2018 23:00:00 +0000 https://www.leica-microsystems.com/19922 Fluorescence Microscopy Widefield Microscopy Bone loss is a serious problem in spaceflight; however, the initial action of microgravity has not been identified. To examine this action, we performed live-imaging of animals during a space mission followed by transcriptome analysis using medaka transgenic lines expressing osteoblast and osteoclastspecific promoter-driven GFP and DsRed. In live-imaging for osteoblasts, the intensity of osterix- or osteocalcin-DsRed fluorescence in pharyngeal bones was significantly enhanced 1 day after launch; and this enhancement continued for 8 or 5 days. https://www.leica-microsystems.com//science-lab/acute-transcriptional-up-regulation-specific-to-osteoblastsosteoclasts-in-medaka-fish-immediately-after-exposure-to-microgravity/ Tue, 13 Feb 2018 23:00:00 +0000 https://www.leica-microsystems.com/19940 Laser Microdissection In this webinar, you will learn: - How to perform isolated proteomics of microscopic regions of interest - How to use this technique with FFPE tissue - How proteomics can help you understand molecular mechanisms in disease https://www.leica-microsystems.com//science-lab/how-to-conduct-localized-proteomics-of-microscopic-regions/ Thu, 08 Feb 2018 23:00:00 +0000 Dr. Eleanor Drummond https://www.leica-microsystems.com/19937 Coherent Raman Scattering (CRS) Heparan Sulfate Proteoglycans act as a cell surface co-receptors for many proteins in which a signal is sent downstream via its cytosolic domain. The role of syndecan-4 proteoglycans in FA assembly was studied in endothelial cells, and single molecule localization super-resolution microscopy showed that Syn4 loss altered the cytoskeleton protein network assembly. Using molecular and cell biology approaches we were able to demonstrate that syndecan-4 plays a pivotal role in endothelial cell behavior. Super-resolution microscopy revealed that syndecan-4 knockdown alters the level and arrangement of essential proteins for focal adhesion, including actin network and a selective decoupling of vinculin from actin, leading to lamellipodia assembly and filopodia protrusions (Figure 1). Furthermore, using Coherent Anti-stokes Raman Scattering (CARS) microscopy, the structure of the tumors formed by such cells in mice was revealed showing its network of vascularization (Figure 2). We propose a model in which syndecan-4 acts as a central mediator: serving as a cell-signaling center in which ECM (fibronectin), cell surface (integrins) and intracellular components (actin and vinculin) come together. https://www.leica-microsystems.com//science-lab/coupling-of-vinculin-to-f-actin-requires-syndecan-4-proteoglycan/ Wed, 07 Feb 2018 23:00:00 +0000 https://www.leica-microsystems.com/19882 Surgical Microscopy In vascular neurosurgery, surgical microscopes are used to provide a magnified and illuminated view of the surgical field. Although surgeons benefit greatly from the superb image quality and optical magnification offered by the microscope, current limitations of the microscope mean that surgeons have to rely on their color acuity to differentiate between different tissue types. https://www.leica-microsystems.com//science-lab/the-future-of-fluorescence-in-vascular-neurosurgery/ Wed, 31 Jan 2018 23:00:00 +0000 PhD George Themelis, MD Robert Ibe https://www.leica-microsystems.com/19812 Widefield Microscopy Live-Cell Imaging Adipose stem cells (ASCs) have recently emerged as a more viable source for clinical applications, compared to bone-marrow mesenchymal stromal cells (BM-MSCs) because of their abundance and easy access. In this study we evaluated the regenerative potency of ASCs compared to BM-MSCs. Furthermore, we compared the dielectric and electro-kinetic properties of both types of cells using a novel Dielectrophoresis (DEP) microfluidic platform based on a printed circuit board (PCB) technology. Our data show that ASCs were more effective than BM-MSCs in promoting neovascularization in an animal model of hind-limb ischemia. https://www.leica-microsystems.com//science-lab/adipose-stem-cells-display-higher-regenerative-capacities-and-more-adaptable-electro-kinetic-properties-compared-to-bone-marrow-derived-mesenchymal-stromal-cells/ Sun, 21 Jan 2018 23:00:00 +0000 https://www.leica-microsystems.com/20478 Super-Resolution A list of the published scientific articles which include work done in the ALMS/MSI Facilities. https://www.leica-microsystems.com//science-lab/cnsi-publication-list/ Sun, 14 Jan 2018 23:00:00 +0000 https://www.leica-microsystems.com/19872 Super-Resolution The 7th Super-Resolution User Club Meeting was held in collaboration with Prof Pavel Hozák , at the Institute of Molecular Genetics of the ASCR in Prague. Keeping the event close to science is one of the founding principles of the event, allowing all participants to network, share and explore exciting new super-resolution and nanoscopy applications. Central to this are the scientific talks given during the meeting, with this cutting-edge microscopy technique as their central theme. A wide selection of topics were covered, prompting interesting discussions during the workshops. https://www.leica-microsystems.com//science-lab/abstracts-of-the-7th-european-super-resolution-user-club-meeting/ Sun, 17 Dec 2017 23:00:00 +0000 Prof. DrSc. Pavel Hozák, Prof. Dr. Torsten Ochsenreiter, Dr. Martin Offterdinger, Dr. Jordi Andilla, Ph.D. Marc van Zandvoort, Prof. Christian Eggeling, Dr. Susan Cox, Dr. Eugene Katrukha, Dr. Jindřiška Fišerová, Dr. Camille Boutin https://www.leica-microsystems.com/19843 Education Wi-Fi Education solutions help teachers to make class more interactive and inspiring. With network capable microscope cameras and the Leica AirLab App learning content gets transferred directly to students’ viewing devices. This promotes teamwork and makes sharing easy in the classroom and beyond. https://www.leica-microsystems.com//science-lab/airteach-software/ Mon, 11 Dec 2017 23:00:00 +0000 Vince Vaccarelli https://www.leica-microsystems.com/19845 Education A challenge for any teacher, especially in the microscopy classroom, is to catch and hold students’ attention. Today the Wi-Fi Education solutions from Leica Microsystems help teachers to make class more interactive and inspiring. With network capable microscope cameras and the Leica AirLab App learning content gets transferred directly to students’ viewing devices. This promotes teamwork and makes sharing easy in the classroom and beyond. https://www.leica-microsystems.com//science-lab/airteach-software/thank-you-for-your-registration-airteach-software-webinar/ Mon, 11 Dec 2017 23:00:00 +0000 Vince Vaccarelli https://www.leica-microsystems.com/19820 Quality Assurance Metallography Obtaining cleanliness results rapidly, accurately, and reliably is a significant advantage for manufacturers and component suppliers. For this reason, Pall Corporation and Leica Microsystems have joined efforts to offer a new, unique workflow solution customizable to your individual needs. Join our free webinar to get a sneak preview on Leica Microsystems' new fully implemented LIBS (Laser Induced Breakdown Spectroscopy) system. This new landmark technology enables you to easily conduct fast and reliable optical and chemical cleanliness analysis with only one system. See for yourself how Leica Microsystems' new software and system solution facilitates and accelerates the identification of nature and source of particle contamination on components. https://www.leica-microsystems.com//science-lab/cleanliness-workflow-from-leica-and-pall/ Wed, 06 Dec 2017 23:00:00 +0000 Dr. Nicol Ecke, Christophe Goasdoué https://www.leica-microsystems.com/19710 Light Sheet Microscopy The TCS SP8 DLS microscope system from Leica Microsystems is an innovative concept which integrates the Light Sheet Microscopy technology into the SP8 confocal platform. Due to its unique optical architecture, samples can be mounted on standard glass bottom petri dishes and require little or no adaptation when compared to conventional mounting procedures. Here, we present a convenient way to prepare specimens quickly for light sheet imaging with the TCS SP8 DLS system. https://www.leica-microsystems.com//science-lab/dls-sample-preparation-using-u-shaped-glass-capillaries-for-sample-mounting/ Wed, 06 Dec 2017 08:45:00 +0000 Dr. Petra Haas https://www.leica-microsystems.com/19773 Fluorescence Microscopy Basics in Microscopy At some point in your research and science career, you will no doubt come across fluorescence microscopy. This ubiquitous technique has transformed the way in which microscopists can image, tag and trace anything from whole organisms to single proteins and beyond. In this article, we will examine what is meant by "fluorescence", the history and basic physics behind its definition, the discovery and application of Green Fluorescent Protein (GFP) and a look at the rapidly expanding field of fluorescent probes including Quantum Dots. https://www.leica-microsystems.com//science-lab/the-fundamentals-and-history-of-fluorescence-and-quantum-dots/ Wed, 15 Nov 2017 16:44:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/24748 https://www.leica-microsystems.com//science-lab/how-to-make-a-fast-z-stack/thank-you-for-your-registration/ Sun, 05 Nov 2017 23:00:00 +0000 https://www.leica-microsystems.com/19766 Multiphoton Microscopy The colorful picture shows colon tumor cells, fluorescently labelled and lineage traced from a multicolor tracer. The gray color codes for the second harmonic generation (SHG) signal from Collagen 1. Lineage traced tumor cells are shown in magenta, blue, green, yellow and red. All channels were recorded with two-photon excitation, using the SP8 DIVE by Leica Microsystems. Sample and image were kindly provided by J. van Rheenen, H. Snippert, Utrecht (the Nederlands,) and I. Steinmetz, Leica Microsystems Mannheim. https://www.leica-microsystems.com//science-lab/multiphoton-microscopy-a-satisfied-wish-list/ Wed, 25 Oct 2017 08:49:00 +0000 Dr. Rolf T. Borlinghaus, Ph.D. Bettina Griesshaber https://www.leica-microsystems.com/19756 EM Sample Preparation Mechanical polishing can be time consuming and frustrating. It can also introduce unwanted artifacts when preparing cross-sectioned samples for electron backscatter diffraction (EBSD) in the scanning electron microscope (SEM) or light microscope investigation. In contrast, ion beam milling can eliminate undesirable artifacts that will hamper your analysis and interpretation. https://www.leica-microsystems.com//science-lab/practical-applications-of-broad-ion-beam-milling/ Thu, 14 Sep 2017 07:50:00 +0000 https://www.leica-microsystems.com/19755 EM Sample Preparation What if you can dissect the cellular dynamics with millisecond precision? What if you can unravel the morphological transformation of a neuron millisecond by millisecond using electron microscopy? https://www.leica-microsystems.com//science-lab/revealing-cellular-dynamics-with-millisecond-precision/ Wed, 13 Sep 2017 07:50:00 +0000 PhD Shigeki Watanabe, PhD Frédéric Leroux https://www.leica-microsystems.com/19722 Digital Microscopy Classical polarized light (compound) microscopes can only be used for prepared samples, because the working distance they offer is insufficient for whole samples. This means that thicker and bigger geological samples have to be sectioned and polished to fit under the limited working distance of a compound microscope. The Leica DVM 6 provides outstanding image quality, extra working distance and allows geologists to work with polished and unpolished samples (micro mounts, fossils, drilled cores …), make 3D reconstructions of their surfaces and enables classical petrographic work as well. https://www.leica-microsystems.com//science-lab/digital-microscopy-in-earth-science/ Wed, 30 Aug 2017 11:51:00 +0000 MSc Michael Doppler https://www.leica-microsystems.com/19749 Digital Microscopy Classical polarized light (compound) microscopes can only be used for prepared samples, because the working distance they offer is insufficient for whole samples. This means that thicker and bigger geological samples have to be sectioned and polished to fit under the limited working distance of a compound microscope. The Leica DVM 6 provides outstanding image quality, extra working distance and allows geologists to work with polished and unpolished samples (micro mounts, fossils, drilled cores …), make 3D reconstructions of their surfaces and enables classical petrographic work as well. https://www.leica-microsystems.com//science-lab/digital-microscopy-in-earth-science/watch-the-webinar-on-demand-digital-microscopy-in-earth-science/ Wed, 30 Aug 2017 11:51:00 +0000 MSc Michael Doppler https://www.leica-microsystems.com/19684 Basics in Microscopy For most microscope applications, there are generally only two sets of optics which are adjusted by the user, namely, the objectives and the eyepieces. Of course, this is assuming that the microscope is already corrected for Koehler Illumination during which the condenser and diaphragms are adjusted. https://www.leica-microsystems.com//science-lab/eyepieces-objectives-and-optical-aberrations/ Mon, 28 Aug 2017 07:36:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19707 Multiphoton Microscopy Leica Microsystems’ 4Tune detector, the key component of the SP8 DIVE Deep In Vivo Explorer, provides spectrally tunable image recording with non-descanning detection. An innovative solution for multiparameter multiphoton microscopy. The colorful image on the right shows multiphoton microscopy of an unstained mouse skin section acquired using the 4Tune detector. The green color codes for autofluorescence of muscle tissue. Red shows second harmonic generation of fibers upon illumination with 900 nm. The blue pattern is generated by third harmonic generation at lipid boundaries from illumination at 1230 nm. https://www.leica-microsystems.com//science-lab/mission-impossible-accomplished-tunable-colors-for-non-descanning-detection/ Wed, 23 Aug 2017 08:49:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Hilmar Gugel https://www.leica-microsystems.com/19639 Basics in Microscopy The technique of Koehler Illumination is one of the most important and fundamental techniques in achieving optimum imaging in any given light microscope set-up. Although it should be routinely used as part of setting up a microscope, many microscopists are put off by thinking that the correct set-up is complex and time consuming and it is therefore still not widely practised. By getting to know the two main components of the microscope which are adjusted in this technique (the diaphragms and sub-stage condenser) in reality, correct set-up should only take a matter of minutes. A correctly aligned microscope can result in greatly improved images of uniform contrast and illumination as well as higher resolution and more detail. In this article, we will look at the history of the technique in addition to how to adjust the components in five easy steps. https://www.leica-microsystems.com//science-lab/koehler-illumination-a-brief-history-and-a-practical-set-up-in-five-easy-steps/ Thu, 17 Aug 2017 07:07:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19686 Live-Cell Imaging Mammalian cell culture is one of the basic pillars of life sciences. Without the ability to grow cells in the lab, the fast progress in disciplines like cell biology, immunology, or cancer research would be unthinkable. This article gives an overview of mammalian cell culture systems. Mainly, they can be categorized according to their morphology, as well as cell type and organization. Moreover, you can find basic information about the correct growth conditions and what kind of microscope you need to watch your cells. https://www.leica-microsystems.com//science-lab/introduction-to-mammalian-cell-culture/ Thu, 10 Aug 2017 10:54:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/19673 Basics in Microscopy To examine specimens at high magnifications using the microscope, there are a number of factors which need to be taken into consideration. These include resolution, numerical aperture (NA), the working distance of objectives and the refractive index of the medium through which the image is collected by the front lens of an objective. In this article, we will briefly look at how using an immersion medium between the coverslip and the objective front lens helps to increase the NA and resolution. https://www.leica-microsystems.com//science-lab/immersion-objectives-using-oil-glycerol-or-water-to-overcome-some-of-the-limits-of-resolution/ Wed, 09 Aug 2017 12:07:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19622 Basics in Microscopy Live-Cell Imaging Heavy metal contamination in water is a serious risk to the public health and other life forms on earth. Current research in nanotechnology is developing new nanosystems and nanomaterials for the fast and efficient removal of pollutants and heavy metals from water. Here, we report graphene oxide-based microbots (GOx-microbots) as active self-propelled systems for the capture, transfer, and removal of a heavy metal (i.e., lead) and its subsequent recovery for recycling purposes. https://www.leica-microsystems.com//science-lab/graphene-based-microbots-for-toxic-heavy-metal-removal-and-recovery-from-water/ Thu, 03 Aug 2017 10:27:00 +0000 https://www.leica-microsystems.com/19635 Fluorescence Microscopy Live-Cell Imaging Stereo Microscopy Widefield Microscopy The zebrafish (Danio rerio) is an emerging vertebrate model organism to study infection. The transparent larva comprises a fully functional innate immune system and enables live imaging of fluorescent immune cells in transgenic animals. Zebrafish infection models have been developed for both the human bacterial pathogen Shigella flexneri and the natural fish bacterial pathogen Mycobacterium marinum. Importantly, whilst S. flexneri causes acute infection and is typically used as an inflammatory paradigm, M. marinum causes a chronic disease similar to tuberculosis in humans. Here, we use real time fluorescence microscopy to image transgenic zebrafish larvae with neutrophils (granulocyte white blood cells) expressing the green fluorescent protein eGFP. https://www.leica-microsystems.com//science-lab/real-time-observation-of-neutrophil-white-blood-cell-recruitment-to-bacterial-infection-in-vivo/ Thu, 27 Jul 2017 11:20:00 +0000 MSc Alexandra R. Willis, MSc Vincenzo Torraca, PhD Serge Mostowy, PhD James DeRose https://www.leica-microsystems.com/19654 Quality Assurance For automotive manufacturers and auto parts suppliers, obtaining component cleanliness results rapidly, accurately, and reproducibly over the entire workflow is a significant advantage. Pall and Leica can make it easier. https://www.leica-microsystems.com//science-lab/free-webinar-on-demand-basics-in-component-cleanliness-analysis/ Wed, 26 Jul 2017 13:31:00 +0000 Dr. Nicol Ecke https://www.leica-microsystems.com/19605 Basics in Microscopy Numerical aperture (abbreviated as ‘NA’) is an important consideration when trying to distinguish detail in a specimen viewed down the microscope. NA is a number without units and is related to the angles of light which are collected by a lens. In calculating NA (see below), the refractive index of a medium is also taken into account and by matching the refractive index of a slide or cell culture container with an immersion medium, then more of the detail of a specimen will be resolved. The way in which light behaves when travelling from one medium to another is also related to NA (and termed ‘refraction’). This article also covers a brief history of refraction and how this concept is a limiting factor in achieving high NA. https://www.leica-microsystems.com//science-lab/collecting-light-the-importance-of-numerical-aperture-in-microscopy/ Wed, 12 Jul 2017 07:46:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19607 EM Sample Preparation Dr. Shigeki Watanabe, principle investigator of the department of Cell Biology at the Johns Hopkins University School of Medicine in Baltimore, held a workshop in Zürich, Switzerland on methods to study synaptic dynamics with millisecond precision. In collaboration with Dr. Andres Käch from the University of Zurich all workshop attendees enjoyed presentations and hands-on sessions on the EM ICE by Leica Microsystems with Light and Electrical Stimulation, revealing the latest developments in brain research. During this workshop Dr. Bernd Sägmüller from Leica Microsystems had the chance for an interview with Dr. Watanabe. https://www.leica-microsystems.com//science-lab/interview-with-dr-shigeki-watanabe-on-research-in-synaptic-membrane-dynamics/ Thu, 06 Jul 2017 23:00:00 +0000 Dr. Bernd Sägmüller, PhD Shigeki Watanabe https://www.leica-microsystems.com/19620 Multiphoton Microscopy Multiphoton Microscopy is one of the current hot topics in life science research. The new Leica TCS SP8 DIVE from Leica Microsystems presents a series of beneficial new innovations, including a freely tunable non-descanning detector and an ingenious beam manipulating device VBE. The variable beam expander offers free tuning of both beam diameter and axial IR-correction for up to four IR beams simultaneously. https://www.leica-microsystems.com//science-lab/laser-beam-shaping-for-multicolor-multiphoton-microscopy/ Thu, 06 Jul 2017 08:24:00 +0000 Dr. Rolf T. Borlinghaus, Manuel Kremer https://www.leica-microsystems.com/19548 Widefield Microscopy Basics in Microscopy One of the most basic microscopy techniques is known as ‘Widefield Microscopy’. It is fundamentally any technique in which the entire specimen of interest is exposed to the light source with the resulting image being viewed either by the observer or a camera (which can also be attached to a computer monitor). https://www.leica-microsystems.com//science-lab/introduction-to-widefield-microscopy/ Thu, 29 Jun 2017 11:31:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19544 Stereo Microscopy Digital Microscopy What do you associate with the word sensor? Perhaps technologies delivering automation in factories and other production sites? That may be what comes to mind, but advances in organic electronics are now driving the rapid development of biological sensors that measure physiological signals when in contact with the skin, organs, and other parts of the body. https://www.leica-microsystems.com//science-lab/coming-to-grips-with-biological-information-through-flexible-organic-electronics-developing-bendable-and-stretchable-biosensors-and-device/ Wed, 28 Jun 2017 10:14:00 +0000 Kuniko Yagura, PhD James DeRose https://www.leica-microsystems.com/19538 Optical Coherence Tomography This article published in the journal OSLI Retina discusses the findings of the latest phase of the DISCOVER study relating specifically to the EnFocus instrasurgical OCT system from Leica. One of the findings of this phase of the study was that “in membrane peeling procedures, intraoperative OCT findings were discordant from the surgeon's initial impression in seven of 20 cases (35%).” (Runkle, Sunil et al, 2017). https://www.leica-microsystems.com//science-lab/microscope-integrated-oct-feasibility-and-utility-with-the-enfocus-system-in-the-discover-study/ Thu, 22 Jun 2017 11:33:00 +0000 https://www.leica-microsystems.com/19239 CRISPR-Cas9 Jennifer Doudna tells the story of how studying the way bacteria fight viral infection turned into a genomic engineering technology that has transformed molecular biology research. In 2013, Doudna and her colleagues developed the CRISPR-Cas9 gene expression system that, when introduced into animal cells, makes site-specific changes to intact genomes. CRISPR-Cas9 is more precise, more efficient, and less expensive than other genome editing tools and, as a result, has facilitated a wide range of studies that were previously unachievable. https://www.leica-microsystems.com//science-lab/video-genome-engineering-with-crispr-cas9/ Fri, 09 Jun 2017 06:50:00 +0000 https://www.leica-microsystems.com/19530 CRISPR-Cas9 Jennifer Doudna tells the story of how studying the way bacteria fight viral infection turned into a genomic engineering technology that has transformed molecular biology research. In 2013, Doudna and her colleagues developed the CRISPR-Cas9 gene expression system that, when introduced into animal cells, makes site-specific changes to intact genomes. CRISPR-Cas9 is more precise, more efficient, and less expensive than other genome editing tools and, as a result, has facilitated a wide range of studies that were previously unachievable. https://www.leica-microsystems.com//science-lab/video-genome-editing-with-crispr-cas9/ Thu, 08 Jun 2017 06:50:00 +0000 https://www.leica-microsystems.com/19342 Super-Resolution The canonical atrial myocyte (AM) is characterized by sparse transverse tubule (TT) invaginations and slow intracellular Ca2+ propagation but exhibits rapid contractile activation that is susceptible to loss of function during hypertrophic remodeling. Here, we have identified a membrane structure and Ca2+-signaling complex that may enhance the speed of atrial contraction independently of phospholamban regulation. This axial couplon was observed in human and mouse atria and is composed of voluminous axial tubules (ATs) with extensive junctions to the sarcoplasmic reticulum (SR) that include ryanodine receptor 2 (RyR2) clusters. In mouse AM, AT structures triggered Ca2+ release from the SR approximately 2 times faster at the AM center than at the surface. https://www.leica-microsystems.com//science-lab/axial-tubule-junctions-control-rapid-calcium-signaling-in-atria/ Wed, 31 May 2017 09:21:00 +0000 https://www.leica-microsystems.com/19234 Basics in Microscopy Optical microscopes are used to magnify objects which are otherwise invisible for the human eye. For this purpose high quality optics is necessary to achieve appropriate resolution. However, besides intentional effects, all optical components have also unwanted intrinsic influence on light, resulting in aberrations. This article highlights optical elements and their physical parameters involved in this process. Based on this, it gives a historical overview of philosophies about how to cope with aberration reduction. Seeing the microscope as a whole system turned out to be beneficial, leading to the harmonization of its constituents for optimal microscopic results. https://www.leica-microsystems.com//science-lab/optimization-of-the-interplay-of-optical-components-for-aberration-free-microscopy/ Wed, 31 May 2017 08:58:00 +0000 Karl-Heinz Schade, Peter Euteneuer, Dipl. Phys. Arnold Müller-Rentz https://www.leica-microsystems.com/19451 Live-Cell Imaging Fluorescence Microscopy Multiphoton Microscopy When operating a confocal microscope, or when discussing features and parameters of such a device, we inescapably mention the pinhole and its diameter. This short introductory document is meant to explain the significance of the pinhole for those, who did not want to spend too much time to dig into theory and details of confocal microscopy but wanted to have an idea about the effect of the pinhole. https://www.leica-microsystems.com//science-lab/five-questions-asked-prof-dr-jacco-van-rheenen-speaks-about-the-most-important-considerations-when-imaging-deep-into-mouse-tissue/ Fri, 19 May 2017 05:47:00 +0000 PhD Jacco van Rheenen, Dr. Bernd Sägmüller https://www.leica-microsystems.com/20002 Grain size plays an essential role in the mechanical properties of materials. Learn how to overcome problems in sample preparation and how to analyze different materials like, steel, aluminium, titanium, copper and ceramics. https://www.leica-microsystems.com//science-lab/analyze-grain-size-on-microstructures-the-way-you-need/watch-the-webinar-analyze-grain-size-on-microstructures-the-way-you-need/ Wed, 17 May 2017 07:50:00 +0000 Dionis Diez https://www.leica-microsystems.com/19444 Quality Assurance Metallography Grain size plays an essential role in the mechanical properties of materials. Learn how to overcome problems in sample preparation and how to analyze different materials like, steel, aluminium, titanium, copper and ceramics. https://www.leica-microsystems.com//science-lab/analyze-grain-size-on-microstructures-the-way-you-need/ Wed, 17 May 2017 07:50:00 +0000 Dionis Diez https://www.leica-microsystems.com/19283 Confocal Microscopy Current fluorescence microscopy employs incident illumination which requires separation of illumination and emission light. The classical device performing this separation is a color-dependent beam splitting mirror which has fixed spectral parameters and transmits the emission usually between 90% and 98% within the designated bands. Transmission is wavelength dependent and also differs by technology, requirements and design. An alternative is the acousto optical beam splitter which has freely tunable reflection notches and transmits the emission on average at 95% between these notches. https://www.leica-microsystems.com//science-lab/primary-beam-splitting-devices-for-confocal-microscopes/ Tue, 16 May 2017 07:42:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/19429 Fluorescence Microscopy The term photomanipulation describes a wide range of techniques that enable the microscopist the transition from passive observer to instigator of events by offering a way of interacting with their sample via targeted illumination. Typically researchers are trying to observe specific processes of interest in order to understand the underlying biological process. Microscopists are often forced to hunt through large populations of cells or acquire hours of time laps footage before they’re able to observe events of interest and in many cases it’s simply not possible to observe certain processes using conventional microscopy techniques alone. Photomanipulation tools enable the microscopist to initiate biological events, precisely adjusting sample labeling, biological activity, local chemical environments and in some instances physically destroy parts of their specimen. https://www.leica-microsystems.com//science-lab/what-is-photomanipulation/ Mon, 15 May 2017 07:59:00 +0000 Dr. Peter Laskey, Dr. Christoph Greb, Dr. Oliver Schlicker https://www.leica-microsystems.com/19440 Digital Microscopy From routine quality assurance to advanced 3D analysis of complex surface structures, there is a digital microscope solution to meet almost any request https://www.leica-microsystems.com//science-lab/quality-with-confidence/ Tue, 09 May 2017 07:50:00 +0000 Clinton Smith, Mario J. Gislao https://www.leica-microsystems.com/6395 Fluorescence Microscopy Fluorescent proteins (FPs) such as GFP, YFP or DsRed are powerful tools to visualize cellular components in living cells. Nevertheless, there are circumstances when classical FPs reach their limits. Watching dedicated, spatially limited protein populations of a certain protein of interest is impossible with common FPs, since they are expressed throughout the entire cell. At this point photoactivatable, photoconvertible and photoswitchable fluorescent proteins enter the stage. The members of this fluorescence toolkit can be activated from a non-fluorescent state, they can change their emission spectrum, or they are even able to be reversibly switched "on and off". With the help of these “optical highlighters”, researchers can track a distinct protein population over time by activating respectively converting their fluorescence with a spatially defined light beam of a given wavelength. https://www.leica-microsystems.com//science-lab/photoactivatable-photoconvertible-and-photoswitchable-fluorescent-proteins/ Thu, 04 May 2017 11:31:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/19158 Optical Coherence Tomography Surgical Microscopy When treating patients with vitreomacular traction (VMT), peeling of the inner limiting membrane (ILM) or an epiretinal membrane (ERM) is often indicated. Successful outcomes depend on precise surgical maneuvers as well as the surgeon’s experience in order to assess whether the entire membrane has been removed, as this is crucial for successful release of traction. Showing different steps of removing an ERM to resolve VMT, the following videos captured with EnFocus intrasurgical OCT demonstrate how visualization of the membrane can impact the surgical process of membrane peeling. https://www.leica-microsystems.com//science-lab/how-intrasurgical-oct-can-impact-the-surgical-approach-to-membrane-peeling/ Thu, 04 May 2017 07:12:00 +0000 Marie Stockton https://www.leica-microsystems.com/19348 Neuroscience Live-Cell Imaging Unlike genetic or pharmacological manipulation, light-controlled proteins respond immediately, can be temporally and spatially triggered, are reversible, and are specific to the protein of interest. However, designing and using light-controllable proteins often requires expertise and specialized equipment. Now, two recently published articles in Science describe generalizable methods for making photo-controllable proteins. https://www.leica-microsystems.com//science-lab/clap-on-clap-off-protein-on-protein-off/ Tue, 02 May 2017 22:27:00 +0000 https://www.leica-microsystems.com/19321 Confocal Microscopy When operating a confocal microscope, or when discussing features and parameters of such a device, we inescapably mention the pinhole and its diameter. This short introductory document is meant to explain the significance of the pinhole for those, who did not want to spend too much time to dig into theory and details of confocal microscopy but wanted to have an idea about the effect of the pinhole. https://www.leica-microsystems.com//science-lab/pinhole-effect-in-confocal-microscopes/ Wed, 26 Apr 2017 10:11:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/19390 Laser Microdissection Seeing is believing, this is what makes microscopes, imaging systems, or box-type imagers so important for researchers in many different fields like pathology research, cancer research, neuroscience, and developmental research. Correlation of phenotype (what you can see) and genotype (genetic background) is highly desired but often biased by whole tissue approaches as no real solution is available to reliably separate the single cells of interest and surrounding tissue. Thus, results of DNA mutation analysis (sequencing), gene expression profiling (quantitative real-time PCR, qPCR, microarray, digital PCR), next generation sequencing (NGS) or mass spectrometry (MS) approaches can give mixed results as no pure starting material for the biomolecular methods of choice is available. https://www.leica-microsystems.com//science-lab/correlation-of-phenotype-and-genotype-microscopic-samples-and-their-biomolecular-backgrounds/ Thu, 20 Apr 2017 07:08:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/19257 Stereo Microscopy Fluorescence Microscopy This report provides information which can help improve the routine work of scientists and technicians performing studies involving surgery on small animals and rodents, i.e. mice, rats, hamsters etc., for developmental biology or medical research. The aim is to help make the work steps efficient and cost-effective, where the employment of microscopes is necessary. It also gives useful hints and details on the various microscopes which can be used in a developmental biology or medical research laboratory where small animal or rodent surgery is exploited. https://www.leica-microsystems.com//science-lab/work-efficiently-in-developmental-biology-and-medical-research-with-stereo-microscopy-rodent-and-small-animal-surgery/ Mon, 27 Mar 2017 08:08:00 +0000 PhD James DeRose, PhD Heinrich Bürgers, MSc Ayyakkannu Ayyanan, MD Cathrin Brisken https://www.leica-microsystems.com/19046 Surgical Microscopy The human skull is an especially delicate area for surgery. To differentiate healthy tissue from tumor cells, or treat an aneurysm for example, the surgeon needs a complete picture of the surgical area. Supplementary information provided by Image Guided Surgery (IGS) or neuro navigation systems, endoscope, and in some cases fluorescence, can support the surgeon’s visualization, insight and intra-surgical orientation. This supplementary visual information is often only viewable on a separate screen, but with image injection technology from Leica it can be overlaid in full high definition (HD) directly into the eyepieces of a surgical microscope. https://www.leica-microsystems.com//science-lab/full-hd-image-injection-provides-a-complete-picture-of-the-surgical-site-directly-in-the-microscope-eyepieces/ Mon, 20 Mar 2017 12:54:00 +0000 https://www.leica-microsystems.com/9474 Surgical Microscopy One of the challenges of neurosurgery is orientation at the surgical site. When resecting tumors, removing arteriovenous malformations, or clipping aneurysms, surgeons often have to work near healthy and functional brain tissue. When resecting the tumor, the challenge is always to spare as much healthy tissue as possible. Neuronavigation technology, also referred to as Image Guided Surgery (IGS) enables surgeons to plan the ideal approach before making a cut and helps to execute that plan by providing intraoperative orientation. https://www.leica-microsystems.com//science-lab/navigating-through-the-brain/ Mon, 20 Mar 2017 12:52:00 +0000 Valentin Elefteriu https://www.leica-microsystems.com/19270 Surgical Microscopy In vascular and neoplastic surgery sylvian dissection is an essential skill. The neurosurgeon needs to recognize the superficial sylvian vein (SSV) in order to safely and accurately achieve a wide opening of the sylvian fissure. A report by Dr. Hisashi Kubota and his colleagues at the Department of Neurosurgery, Kindai University Faculty of Medicine, Osaka, Japan, discusses the benefits of indocyanine green (ICG) videoanigiography to demarcate the SSV at the beginning of a sylvian dissection, particularly in patients with subarachnoid hemorrhage. Observation of ICG was enabled by an FL800 vascular fluorescence module from Leica in combination with an M525 OH4 neurosurgical microscope. https://www.leica-microsystems.com//science-lab/sylvian-dissection-with-icg-viewed-with-fl800-fluorescence/ Wed, 15 Mar 2017 13:37:00 +0000 Marie Stockton https://www.leica-microsystems.com/18094 EM Sample Preparation Species: red blood worm (midge larva) Critical point drying of midge larvae with subsequent X-ray micro-computed tomography (micro-CT) to reconstruct the inner anatomy. https://www.leica-microsystems.com//science-lab/micro-ct-of-insect-larva-protocol/ Tue, 14 Mar 2017 07:14:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/19260 Basics in Microscopy Fluorescence Microscopy Since the middle of the last century, fluorescence microscopy developed into a bio scientific tool with one of the biggest impacts on our understanding of life. Watching cells and proteins with the help of fluorescence molecules is a standard method in nearly every life science discipline today. This broad application range goes back to the technical work of some researchers who wanted to improve and simplify fluorescence microscopic labor. One person who was involved in that development was the Dutch medic Johann Sebastiaan Ploem. https://www.leica-microsystems.com//science-lab/milestones-in-incident-light-fluorescence-microscopy/ Mon, 06 Mar 2017 08:39:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/19232 Fluorescence Microscopy CRISPR-Cas9 Francisco Mojica was not the first to see CRISPR, but he was probably the first to be smitten by it. He remembers the day in 1992 when he got his first glimpse of the microbial immune system that would launch a biotechnology revolution. He was reviewing genome-sequence data from the salt-loving microbe Haloferax mediterranei and noticed 14 unusual DNA sequences, each 30 bases long. https://www.leica-microsystems.com//science-lab/five-big-mysteries-about-crisprs-origins/ Mon, 20 Feb 2017 08:49:00 +0000 https://www.leica-microsystems.com/19109 Confocal Microscopy While distinct stages of natural killer (NK) cell development have been defined, the molecular interactions that shape human NK cell maturation are poorly understood. Here we define intercellular interactions between developing NK cells and stromal cells which, through contact-dependent mechanisms, promote the generation of mature, functional human NK cells from CD34+ precursors. We show that developing NK cells undergo unique, developmental stage-specific sustained and transient interactions with developmentally supportive stromal cells, and that the relative motility of NK cells increases as they move through development in vitro and ex vivo. https://www.leica-microsystems.com//science-lab/human-nk-cell-development-requires-cd56-mediated-motility-and-formation-of-the-developmental-synapse/ Wed, 15 Feb 2017 07:36:00 +0000 Ph.D. Emily M. Mace https://www.leica-microsystems.com/19107 Fluorescence Microscopy Confocal Microscopy Long distance transport in plants occurs in sieve tubes of the phloem. The pressure flow hypothesis introduced by Ernst Münch in 1930 describes a mechanism of osmotically generated pressure differentials that are supposed to drive the movement of sugars and other solutes in the phloem, but this hypothesis has long faced major challenges. The key issue is whether the conductance of sieve tubes, including sieve plate pores, is sufficient to allow pressure flow. We show that with increasing distance between source and sink, sieve tube conductivity and turgor increases dramatically in Ipomoea nil. Our results provide strong support for the Münch hypothesis, while providing new tools for the investigation of one of the least understood plant tissues. https://www.leica-microsystems.com//science-lab/testing-the-muench-hypothesis-of-long-distance-phloem-transport-in-plants/ Tue, 14 Feb 2017 09:08:00 +0000 https://www.leica-microsystems.com/15098 Basics in Microscopy This lecture describes the famous experiments of Ernst Abbe which showed how diffraction of light by a specimen (and interference with the illuminating light) gives rise to an image and how collection of diffracted light defines the resolution of the microscope. These concepts are demonstrated by using a diffraction grating as a specimen and visualizing and comparing the diffraction pattern in the back focal plane as well as the image in the image plane. https://www.leica-microsystems.com//science-lab/video-talk-by-kurt-thorn-the-abbe-diffraction-experiment/ Mon, 13 Feb 2017 12:12:00 +0000 PhD Kurt Thorn https://www.leica-microsystems.com/18921 Super-Resolution Hemidesmosomes have been extensively studied by immunofluorescence microscopy, but due to its limited resolution, their precise organization remained poorly understood. We studied hemidesmosome organization in cultured keratinocytes by 2- and 3-color super-resolution microscopy. We observed that in the cell periphery, nascent hemidesmosomes are associated with individual keratin filaments and that β4 is distributed along rather than under keratin filaments. By applying innovative methods to quantify molecular distances, we demonstrate that the hemidesmosomal plaque protein plectin interacts simultaneously and asymmetrically with β4 and keratin. https://www.leica-microsystems.com//science-lab/the-molecular-architecture-of-hemidesmosomes-as-revealed-by-super-resolution-microscopy/ Fri, 10 Feb 2017 09:11:00 +0000 MSc Leila Nahidiazar https://www.leica-microsystems.com/19216 Ergonomics Stereo Microscopy Microscopes are tools that affect those who work with them every day. They can be highly demanding for the human body, requiring concentration and a lot of steady activity from many of our muscles. In this interview, Clinton Smith, Senior Product Manager at Leica Microsystems, talks about how to relieve possible tension and strain and how to create ergonomic workplaces to help microscope users work in comfort and how to increase productivity. https://www.leica-microsystems.com//science-lab/ergonomics/how-to-turn-microscope-workplaces-ergonomic/ Thu, 09 Feb 2017 13:08:00 +0000 Claudia Müller https://www.leica-microsystems.com/18172 Multiphoton Microscopy Fluorescence Microscopy Chronic stress induces signalling from the sympathetic nervous system (SNS) and drives cancer progression, although the pathways of tumour cell dissemination are unclear. Here we show that chronic stress restructures lymphatic networks within and around tumours to provide pathways for tumour cell escape. We show that VEGFC derived from tumour cells is required for stress to induce lymphatic remodelling and that this depends on COX2 inflammatory signalling from macrophages. Pharmacological inhibition of SNS signalling blocks the effect of chronic stress on lymphatic remodelling in vivo and reduces lymphatic metastasis in preclinical cancer models and in patients with breast cancer. https://www.leica-microsystems.com//science-lab/chronic-stress-in-mice-remodels-lymph-vasculature-to-promote-tumour-cell-dissemination/ Tue, 07 Feb 2017 09:52:00 +0000 https://www.leica-microsystems.com/18098 Super-Resolution The cytoskeleton is an early attribute of cellular life, and its main components are composed of conserved proteins. The actin cytoskeleton has a direct impact on the control of cell size in animal cells, but its mechanistic contribution to cellular growth in plants remains largely elusive. Here, we reveal a role of actin in regulating cell size in plants. The actin cytoskeleton shows proximity to vacuoles, and the phytohormone auxin not only controls the organization of actin filaments but also impacts vacuolar morphogenesis in an actin-dependent manner. https://www.leica-microsystems.com//science-lab/actin-dependent-vacuolar-occupancy-of-the-cell-determines-auxin-induced-growth-repression/ Fri, 03 Feb 2017 08:14:00 +0000 https://www.leica-microsystems.com/18221 EM Sample Preparation Application Note - Ion milling is a perfect alternative for chemical etching, especially for polycrystalline metals, such as copper. Ion milling can be used to increase the contrast of the grain structure and their interfaces. In contrast to chemical etching the milling process is clean, safe and easy to operate. Ion energy and milling time depend on the milling rate of the metal. https://www.leica-microsystems.com//science-lab/contrast-enhancement-of-polycrystalline-metals-sample-preparation-for-sem/ Wed, 01 Feb 2017 10:33:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/19182 EM Sample Preparation Human peripheral blood contains RNA in cells and in extracellular membrane vesicles, microvesicles and exosomes, as well as in cell-free ribonucleoproteins. Circulating mRNAs and noncoding RNAs, being internalized, possess the ability to modulate vital processes in recipient cells. In this study, with SOLiD sequencing technology, we performed identification, classification, and quantification of RNAs from blood fractions: cells, plasma, plasma vesicles pelleted at 16,000 https://www.leica-microsystems.com//science-lab/variety-of-rnas-in-peripheral-blood-cells-plasma-and-plasma-fractions/ Mon, 30 Jan 2017 10:22:00 +0000 https://www.leica-microsystems.com/19176 EM Sample Preparation Application Note for Leica EM ACE900 - Drosophila larvae were sandwiched between two 3 mm aluminum slit carriers with the 100 μm cavities facing each other and high-pressure frozen with a Leica EM HPM100. No ethanol as synchronization media was used, 1-hexadecene was used as filler. The wholes of the slit carriers were filled with filter tips dipped in 1-hexadecene to keep the carrier sandwich complete after freezing. https://www.leica-microsystems.com//science-lab/drosophila-larvae-sample-preparation-for-cryo-sem/ Mon, 30 Jan 2017 09:50:00 +0000 Dr. Andres Kaech, Prof. Damian Brunner https://www.leica-microsystems.com/19179 EM Sample Preparation Application Note for Leica EM ACE900 - A 100 mesh copper grid (12 um thickness) was dipped into a concentrated Giardia suspension and sandwiched between two flat 3 mm aluminum specimen carriers with scratched surfaces. Subsequently, the sandwich was transferred to the widened hole of a middle plate (3.1 mm diameter). A 50 um spacer ring was added on top and the specimen immediately frozen with an HPM100 high-pressure freezing machine without using alcohol as synchronization fluid. https://www.leica-microsystems.com//science-lab/giardia-lamblia-sample-preparation-for-cryo-sem/ Mon, 30 Jan 2017 09:35:00 +0000 Dr. Andres Kaech, Joe Paulin Zumthor https://www.leica-microsystems.com/19111 Confocal Microscopy Light Sheet Microscopy Multiphoton Microscopy Neuroscience Biological specimens are intrinsically three dimensional; however because of the obscuring effects of light scatter, imaging deep into a tissue volume is problematic. Although efforts to eliminate the scatter by “clearing” the tissue have been ongoing for over a century, there have been a large number of recent innovations. This review introduces the physical basis for light-scatter in tissue, describes the mechanisms underlying various clearing techniques, and discusses several of the major advances in light microscopy for imaging cleared tissue. https://www.leica-microsystems.com//science-lab/clarifying-tissue-clearing/ Thu, 26 Jan 2017 14:39:00 +0000 PhD Douglas Richardson, Dr. Jeff Lichtman https://www.leica-microsystems.com/18958 Super-Resolution Application example of HyVolution Super-Resolution - Atg9 is a multispanning transmembrane protein that is required for autophagosome formation. During autophagy, vesicles containing Atg9 are generated through an unknown mechanism and delivered to the autophagosome formation sites. We have previously reported that Atg9-containing membranes undergo continuous tubulation and fission during nutrient starvation in a manner dependent on the curvature-inducing protein Bif-1/Sh3glb1. Here, we identify Dynamin 2 (DNM2) as a Bif-1-interacting protein that mediates the fission of Atg9-containing membranes during autophagy. https://www.leica-microsystems.com//science-lab/the-bif-1-dynamin-2-membrane-fission-machinery-regulates-atg9-containing-vesicle-generation-at-the-rab11-positive-reservoirs/ Wed, 25 Jan 2017 13:00:00 +0000 https://www.leica-microsystems.com/18156 EM Sample Preparation Cathodoluminescence can be used to achieve spectra and high resolution images of impurity and structural defects in semicondoctors, minerals and insulating materials. This application note explains how to prepare a perfect sample surface for carhodoluminescence and how to use ion beam slope cutting to prepare the sample surface free of any preparation artefacts. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-cadmiumsulphide-cds-for-cathodoluminescence/ Tue, 24 Jan 2017 18:12:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18207 EM Sample Preparation Application Note for Leica EM AMW - Plants (5-years old) were grown in pots filled with soil and kept in greenhouse conditions. Five weeks before harvesting the plants were transferred into growth chambers and cultivated at a temperature of 20°C during daytime and 12°C overnight. The relative humidity was set at 60% and the photoactive radiation was 500 μmol m-2 s-1 during daytime. Sample preparation for transmission electron microscopy (TEM) was performed in order to develop a standard protocol that would reduce sample preparation time for TEM-investigations. Therefore the overall and fine structure of leaf cells prepared with the Leica EM AMW were compared with leaf cells that were prepared with a conventional fixation protocol at room temperature. https://www.leica-microsystems.com//science-lab/picea-abies-l-karst-sample-preparation-for-tem/ Mon, 23 Jan 2017 19:24:00 +0000 Prof. Bernd Zechmann, Prof. Günther Zellnig https://www.leica-microsystems.com/18196 EM Sample Preparation Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating diffraction patterns (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphological studies. The information depth is just a few nm, therefore good sample preparation is very important to avoid damages. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-copper-for-electron-backscattered-diffraction-ebsd/ Thu, 19 Jan 2017 16:37:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18247 EM Sample Preparation Application Note for Leica EM RES102 - Ceramic membrane filters with pore sizes down to a few nanometres must be investigated in cross-section with regard to the structure of the pores. The smallest pores are of special interest. In most cases, conventional grinding methods cannot be used for such problems, as the pore structure would be distorted. This applies in particular to the pores in the nanometre range. https://www.leica-microsystems.com//science-lab/porous-ceramics-sample-preparation-for-sem/ Tue, 17 Jan 2017 07:32:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18319 EM Sample Preparation Application Note for Leica EM HPM100 - Leaves were immersed in hexadecene and placed under a gentle (0.3 bar) vacuum for 10 minutes to evacuate the internal air spaces. The leaves were then trimmed to fit the carriers and placed in the 200 μm side of a 6 mm Type A specimen carrier. Free space was filled with additional hexadecene after which a 6 mm Type B specimen carrier was placed on top with the flat side down. https://www.leica-microsystems.com//science-lab/maple-acer-saccharum-leaves-high-pressure-freezing-and-freeze-substitution-for-tem/ Mon, 16 Jan 2017 17:08:00 +0000 Dr. Kim Rensing https://www.leica-microsystems.com/15854 Digital Microscopy Quality Assurance Surface Metrology State-of-the-art digital microscopes utilizing a versatile illumination system capable of achieving multiple contrast methods, such as the Leica DVM6, are very useful for inspection, quality control, and failure analysis. These contrast methods allow flaws or defects on the surface of a product or component to be more easily and rapidly detected. https://www.leica-microsystems.com//science-lab/digital-microscopy-with-versatile-illumination-and-various-contrast-methods-for-more-efficient-inspection-and-quality-control/ Mon, 16 Jan 2017 10:53:00 +0000 PhD James DeRose, Georg Schlaffer https://www.leica-microsystems.com/18937 Super-Resolution Neuroscience Application example of HyVolution Super-Resolution - Mutations in the CLN1 gene that encodes Palmitoyl protein thioesterase 1 (PPT1) or CLN1, cause Infantile NCL (INCL, MIM#256730). PPT1 removes long fatty acid chains such as palmitate from modified cysteine residues of proteins. The data shown here result from isolated protein complexes from PPT1-expressing SH-SY5Y stable cells that were subjected to single step affinity purification coupled to mass spectrometry (AP-MS). Prior to the MS analysis, we utilised a modified filter-aided sample preparation (FASP) protocol. Based on label free quantitative analysis of the data by SAINT, 23 PPT1 interacting partners (IP) were identified. https://www.leica-microsystems.com//science-lab/quantitative-analysis-of-ppt1-interactome-in-human-neuroblastoma-cells/ Fri, 13 Jan 2017 16:23:00 +0000 https://www.leica-microsystems.com/15140 Stereo Microscopy The female Aedes aegypti salivary gland plays a pivotal role in bloodmeal acquisition and reproduction, and thereby dengue virus (DENV) transmission. It produces numerous immune factors, as well as immune-modulatory, vasodilatory, and anti-coagulant molecules that facilitate blood-feeding. To assess the impact of DENV infection on salivary gland physiology and function, we performed a comparative genome-wide microarray analysis of the naïve and DENV infection-responsive A. aegypti salivary gland transcriptomes. https://www.leica-microsystems.com//science-lab/dengue-virus-infection-of-the-aedes-aegypti-salivary-gland-and-chemosensory-apparatus-induces-genes-that-modulate-infection-and-blood-feeding-behavior/ Thu, 12 Jan 2017 12:29:00 +0000 https://www.leica-microsystems.com/18249 EM Sample Preparation Application Note for Leica EM RES102 - Sometimes it is necessary to remove surface layers to gain access to the real surface structure. That can be a native oxide, or layers coming from the preparation process itself, like re-deposition. Depending on the layers thickness and the energy used for the cleaning process, it takes between a few seconds and half an hour. The energy depends on the milling rate of the material. https://www.leica-microsystems.com//science-lab/removal-of-surface-layers-sample-preparation-for-sem-and-tem/ Wed, 11 Jan 2017 10:10:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18994 Fluorescence Microscopy Live-Cell Imaging Widefield Microscopy In the course of chronic inflammation certain body areas are recurrently inflamed. This goes along with many human diseases. With the help of widefield light microscopy, the underlying processes can be examined from a cellular level to whole organisms. This article presents several widefield microscopy applications such as immunofluorescence, live-cell imaging, histology, and ratiometric analysis to get insight into the development of chronic inflammation, the related diseases, and their treatment. https://www.leica-microsystems.com//science-lab/chronic-inflammation-under-the-microscope/ Mon, 09 Jan 2017 17:42:00 +0000 M.Sc. Jan Neumann, M.Sc. Anne Scherhag, Susanne Otten, Ph.D. Fangxia Shen, Anna Lena Leifke, Dr. Udo Birk, Dr. Christoph Greb, Dr. Kurt Lucas, Prof. Dr. Dr. Christoph Cremer https://www.leica-microsystems.com/18278 Super-Resolution Live-Cell Imaging Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid–protein interactions and the traditional lipid ‘raft’ theory. https://www.leica-microsystems.com//science-lab/super-resolution-optical-microscopy-of-lipid-plasma-membrane-dynamics/ Fri, 23 Dec 2016 14:16:00 +0000 Prof. Christian Eggeling https://www.leica-microsystems.com/18354 EM Sample Preparation Application Note for Leica EM TIC020, Leica EM TIC 3X - Cross section of a complete solar cell. https://www.leica-microsystems.com//science-lab/cross-section-of-solar-cells/ Fri, 23 Dec 2016 06:47:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18362 EM Sample Preparation Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating a diffraction pattern (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphology studies. The information depth is just a few nm. Therefore good sample preparation is very important to avoid any damage. This is very difficult in case of multilayer system with big differences in hardness. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-a-multilayer-system-preparation-of-a-perfect-sample-surface-for-ebsd/ Thu, 22 Dec 2016 20:48:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18317 EM Sample Preparation Application Note for Leica EM HPM100 - Hep-2 cells infected with Chlamydia pneumoniae were cultured on carbon-coated 6 mm Sapphire discs. Cells were high-pressure frozen in an EM HPM100 using the 6 mm CLEM middle plate with following setup: Sapphire disc with cells, spacer 200 μm, bare Sapphire disc, 2 spacers 200 μm. Ethanol was used as a synchronization fluid to transfer pressure at room temperature prior to cooling. https://www.leica-microsystems.com//science-lab/high-pressure-freezing-and-freeze-substitution-of-hep-2-cells-infected-with-chlamydia-pneumoniae/ Tue, 20 Dec 2016 11:03:00 +0000 Dr. Andres Kaech https://www.leica-microsystems.com/18243 EM Sample Preparation Application Note for Leica EM RES102 - The multi-layer system to be prepared in cross-section consists of a Si substrate, a TiN layer with a thickness of a few nm and a 500 nm W layer. All these components have extreme differences in their hardness, their atomic weight and in their sputter rates. A preparation of this kind of samples with sample rotation would lead to a wall overlying the area of the layers. https://www.leica-microsystems.com//science-lab/multilayer-systems-with-widely-different-sputter-rates-sample-preparation-for-tem/ Fri, 16 Dec 2016 16:57:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18008 CLEM Fluorescence Microscopy EM Sample Preparation Correlative light and electron microscopy is an imaging technique that enables identification and targeting of fluorescently tagged structures with subsequent imaging at near-to-nanometer resolution. We established a novel correlative cryo-fluorescence microscopy and cryo-scanning electron microscopy workflow, which enables imaging of the studied object of interest very close to its natural state, devoid of artifacts caused for instance by slow chemical fixation. This system was tested by investigating the interaction of the zoonotic bacterium Borrelia burgdorferi with two mammalian cell lines of neural origin in order to broaden our knowledge about the cell-association mechanisms that precedes the entry of the bacteria into the cell. https://www.leica-microsystems.com//science-lab/correlative-cryo-fluorescence-and-cryo-scanning-electron-microscopy-as-a-straightforward-tool-to-study-host-pathogen-interactions/ Tue, 13 Dec 2016 07:58:00 +0000 Martin Strnad https://www.leica-microsystems.com/18225 EM Sample Preparation Application Note for Leica EM RES102 - Previous studies showed that surface accumulation of In occurs when InP was milled in a conventional way with Ar ions. The consequence is In islands on the sample surface. This leads to low quality of TEM samples. To remove these islands, reactive ion milling with iodine ions (RIBE / CAIBE) can be used. This method has the disadvantage of polluting the ion guns and the vacuum system of the ion milling device and leads to chemical reactions with the sample material. To avoid these problems we prepared these samples very gently with low energy Ar ions. https://www.leica-microsystems.com//science-lab/in-containing-compound-semiconductors-sample-preparation-for-tem/ Mon, 12 Dec 2016 10:13:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18925 Super-Resolution Confocal Microscopy Application example of HyVolution Super-Resolution - Accurate distance measurement in 3D confocal microscopy is important for quantitative analysis, volume visualization and image restoration. However, axial distances can be distorted by both the point spread function (PSF) and by a refractive-index mismatch between the sample and immersion liquid, which are difficult to separate. Additionally, accurate calibration of the axial distances in confocal microscopy remains cumbersome, although several high-end methods exist. In this paper we present two methods to calibrate axial distances in 3D confocal microscopy that are both accurate and easily implemented. https://www.leica-microsystems.com//science-lab/methods-to-calibrate-and-scale-axial-distances-in-confocal-microscopy-as-a-function-of-refractive-index/ Fri, 09 Dec 2016 08:29:00 +0000 https://www.leica-microsystems.com/19036 Basics in Microscopy Live-Cell Imaging Fluorescence Microscopy An optical microscope is often one of the central devices in a life-science research lab. It can be used for various applications which shed light on many scientific questions. Thereby the configuration and features of the microscope are crucial for its application coverage, ranging from brightfield through fluorescence microscopy to live-cell imaging. This article provides a brief overview of the relevant microscope features and wraps up the key questions one should consider when selecting a research microscope. https://www.leica-microsystems.com//science-lab/factors-to-consider-when-selecting-a-research-microscope/ Thu, 08 Dec 2016 15:22:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/18558 Laser Microdissection Application example for laser microdissection (LMD) - MicroRNAs (miRNAs) play important roles in regulating biological processes at the post-transcriptional level. Deregulation of miRNAs has been observed in cancer, and miRNAs are being investigated as potential biomarkers regarding diagnosis, prognosis and prediction in cancer management. Real-time quantitative polymerase chain reaction (RT-qPCR) is commonly used, when measuring miRNA expression. Appropriate normalisation of RT-qPCR data is important to ensure reliable results. The aim of the present study was to identify stably expressed miRNAs applicable as normaliser candidates in future studies of miRNA expression in rectal cancer. https://www.leica-microsystems.com//science-lab/microrna-expression-profiling-to-identify-and-validate-reference-genes-for-the-relative-quantification-of-microrna-in-rectal-cancer/ Wed, 07 Dec 2016 14:44:00 +0000 https://www.leica-microsystems.com/18969 Surgical Microscopy With its high magnification and coaxial illumination, a dental microscope provides enhanced visualization that supports diagnosis and treatment. Its documentation capabilities also play a major role in a modern dental practice by supporting patient consultation. We interviewed Dr. Thorsten Wegner, an endodontist based in Stadthagen, Germany, to find out why he uses the microscope in all aspects of his dental practice. https://www.leica-microsystems.com//science-lab/good-reasons-why-to-use-a-dental-microscope-interview-with-endodontist-dr-thorsten-wegner/ Tue, 06 Dec 2016 11:26:00 +0000 Dr. med. dent. Thorsten Wegner https://www.leica-microsystems.com/19003 EM Sample Preparation Application Note fo Leica EM ACE600 - A solid understanding of polymer property-structure relationships is critical to improve and shorten development routes to new products. A direct way to determine correlations between structure and mechanical properties is provided by electron microscopy. Electron microscopy techniques have an important advantage over other methods, as they can provide local information at high spatial resolution. However, a major problem with polymers is their inherent lack of contrast. https://www.leica-microsystems.com//science-lab/carbon-coating-for-polymeric-materials/ Mon, 05 Dec 2016 09:22:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18991 Basics in Microscopy In microscopy, the term ‘resolution’ is used to describe the ability of a microscope to distinguish detail. In other words, this is the minimum distance at which two distinct points of a specimen can still be seen - either by the observer or the microscope camera - as separate entities. The resolution of a microscope is intrinsically linked to the numerical aperture (NA) of the optical components as well as the wavelength of light which is used to examine a specimen. In addition, we have to consider the limit of diffraction which was first described in 1873 by Ernst Abbe. This article covers some of the history behind these concepts as well as explaining each using relatively simple terminology. https://www.leica-microsystems.com//science-lab/microscope-resolution-concepts-factors-and-calculation/ Fri, 02 Dec 2016 14:09:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/19022 Optical Coherence Tomography Surgical Microscopy Optical Coherence Tomography (OCT) is a non-invasive, non-contact imaging modality used to visualize and monitor changes to the morphology of biological tissue. OCT employs low-coherence interferometry to create cross-sectional images that reveal sub-surface details of the tissues of interest. In the most common ophthalmic applications OCT systems use near-infrared light to generate high-resolution, volumetric images of tissue microstructures including the cornea, iris, crystalline lens, vitreous and retina. These images can enhance insight into pathological conditions such as glaucoma, age-related macular degeneration or diabetic retinopathy. https://www.leica-microsystems.com//science-lab/what-is-oct-and-how-can-it-help-ophthalmologists-acquire-high-resolution-information-on-ocular-tissue/ Thu, 01 Dec 2016 14:44:00 +0000 George Wildeman, PhD Hafeez Dhalla https://www.leica-microsystems.com/18978 Optical Coherence Tomography Surgical Microscopy Replacing diseased Descemet’s membrane with a donor graft is an especially delicate procedure. Accurate positioning and full adherence to the stroma are crucial for optimal patient outcomes. These videos illustrate how intrasurgical OCT can provide additional information during each stage of a DMEK procedure. https://www.leica-microsystems.com//science-lab/how-intrasurgical-oct-can-provide-additional-information-during-ophthalmic-dmek-procedures/ Thu, 01 Dec 2016 14:05:00 +0000 Marie Stockton https://www.leica-microsystems.com/18935 Fluorescence Microscopy Confocal Microscopy Using Arabidopsis (Arabidopsis thaliana) seedlings, we identified a range of small fluorescent probes that entered the translocation stream and were unloaded at the root tip. These probes had absorbance/emission maxima ranging from 367/454 to 546/576 nm and represent a versatile toolbox for studying phloem transport. Of the probes that we tested, naturally occurring fluorescent coumarin glucosides (esculin and fraxin) were phloem loaded and transported in oocytes by the sucrose transporter, AtSUC2. Arabidopsis plants in which AtSUC2 was replaced with barley (Hordeum vulgare) sucrose transporter (HvSUT1), which does not transport esculin in oocytes, failed to load esculin into the phloem. https://www.leica-microsystems.com//science-lab/multispectral-phloem-mobile-probes-properties-and-applications/ Tue, 29 Nov 2016 16:44:00 +0000 https://www.leica-microsystems.com/18927 Super-Resolution Application example of HyVolution Super-Resolution - Bacteria belonging to the genus Cronobacter spp. have been recognized as causative agents of life-threatening systemic infections, primarily in premature, low-birth weight and/or immune-compromised neonates. Knowledge remains scarce regarding the underlying molecular mechanisms of disease development. In this study, we evaluated the use of a zebrafish model to study the pathogenesis of Cronobacter turicensis LMG 23827T, a clinical isolate responsible for two fatal sepsis cases in neonates. https://www.leica-microsystems.com//science-lab/evaluation-of-zebrafish-as-a-model-to-study-the-pathogenesis-of-the-opportunistic-pathogen-cronobacter-turicensis/ Fri, 25 Nov 2016 14:43:00 +0000 https://www.leica-microsystems.com/18983 EM Sample Preparation Application Note for Leica EM ACE600 - Carbon support films are routinely used for high resolution TEM. Thickness is one of the main criteria to assess its usefulness for a particular experiment. Within that respect graphene (oxide) layers are frequently used. However, charge dissipation and mechanical stability towards high probe currents and high voltages, including long term acquisition protocols are equally important. https://www.leica-microsystems.com//science-lab/high-resolution-carbon-coating-how-much-carbon-is-too-much/ Wed, 23 Nov 2016 11:56:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18976 EM Sample Preparation Application Note for Leica EM ACE600 - Focused ion beam (FIB) technology has become an indispensable tool for site-specific TEM sample preparation. It allows to extract electron transparent specimens with nanometer precision using a focused Ga+ ion beam. https://www.leica-microsystems.com//science-lab/each-atom-counts-protect-your-samples-prior-to-fib-processing/ Wed, 23 Nov 2016 11:43:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18932 Education Selecting educational microscopes is not an easy task for teachers. The microscopes must stand up to daily use by not always careful hands, be constantly up and running, and fit the budget requirements. Especially with student microscopes, practical aspects play a role: Size, weight, cabling, and design are important in their daily use – and this should be taken into consideration even before the equipment and accessories of the microscopes enter the decision process. If chosen carefully, educational microscopes will open windows to a cosmos of minute detail which delight young minds in schools and universities – and ideally keeps them fascinated enough to make science their profession. https://www.leica-microsystems.com//science-lab/factors-to-consider-when-selecting-student-microscopes-for-schools-and-universities/ Tue, 22 Nov 2016 09:00:00 +0000 Claudia Müller https://www.leica-microsystems.com/18312 EM Sample Preparation Application Note for Leica EM HPM100 - Hand creams having different water contents were applied into the 100 μm cavities of two 3 mm type A sample carriers which were then closed cream sides inwards. The sample assembly was high pressure frozen with a Leica EM HPM100 and moved to a cooled Leica EM VCT100 loading station. https://www.leica-microsystems.com//science-lab/commercially-available-hand-creams-sample-preparation-for-cryo-sem/ Fri, 18 Nov 2016 17:22:00 +0000 Dr. Kim Rensing https://www.leica-microsystems.com/18255 EM Sample Preparation Application Note for Leica EM RES102 - The cross-sectional preparation of structured semiconductor materials requires a very thorough mechanical pre-preparation. In doing this, it must be ensured that the structure of interest should be located as close to the centre of the sample as possible. As the sample will be ion milled from both sides, a specific preparation of the structure is necessary in most cases, which means that you must thin these structures from both sides. https://www.leica-microsystems.com//science-lab/shallow-trench-isolation-structures-sample-preparation-for-tem/ Fri, 18 Nov 2016 16:50:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18110 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of tobacco leafs with subsequent platinum coating and SEM analysis. https://www.leica-microsystems.com//science-lab/tobacco-leaf-critical-point-drying-protocol-for-sem/ Fri, 18 Nov 2016 16:21:00 +0000 Dr. Martin W. Goldberg, M.Sc. Christine Richardson https://www.leica-microsystems.com/18885 Fluorescence Microscopy CRISPR-Cas9 The CRISPR/Cas9 system is one of several different bacterial systems for defense against viral attacks. It consists of two main components. One is a small piece of RNA which binds to the viral target sequence via Watson-Crick base pairing. It serves as a marker for the foreign nucleic acid. The second component is the Cas9 protein. It binds to the marked sequence and cuts it due to its nuclease activity. Because the base pairing RNA can be synthesized easily and then used to determine a target region, researchers have utilized this system in the laboratory for genome editing. https://www.leica-microsystems.com//science-lab/gene-editing-with-crisprcas9-breakthrough-in-genome-engineering/ Thu, 17 Nov 2016 18:54:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/18245 EM Sample Preparation Application Note for Leica EM RES102 - A coated paper sample has been prepared with ion beam slope cutting in order to test the procedure with regard to its applicability. With the use of ion beam slope cutting a cross section of paper could be prepared. On the basis of this sample processing, it was possible to show the largely unaffected original structure of the thermally-sensitive paper in the scanning electron microscope. https://www.leica-microsystems.com//science-lab/paper-samples-sample-preparation-for-sem/ Wed, 16 Nov 2016 14:01:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18241 EM Sample Preparation Application Note for Leica EM RES102 - Most metal films already have a thickness that requires no further mechanical pre-preparation. Frequently, however, they are also domed, which can lead to undefined milling angles. This is a disadvantage, particularly for films that contain inclusions, and that therefore mostly require very flat milling angles. Metal sheets are thicker than 100 µm. Mechanical pre-preparation is necessary to obtain an acceptable initial thickness and a good surface quality for ion milling. https://www.leica-microsystems.com//science-lab/metal-films-and-sheets-sample-preparation-for-tem/ Wed, 16 Nov 2016 13:32:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18106 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of nematode Eubostrichus dianae to detect the ectosymbiotic bacteria layer with subsequent gold coating ans SEM analysis. https://www.leica-microsystems.com//science-lab/nematode-e-dianae-critical-point-drying-protocol-for-sem/ Wed, 16 Nov 2016 11:05:00 +0000 Mag. Nikolaus Leisch https://www.leica-microsystems.com/18952 Super-Resolution Live-Cell Imaging Application example of HyVolution Super-Resolution - Atherosclerosis occurs preferentially in arterial regions exposed to disturbed blood flow. Targeting these pro-atherogenic regions is a potential anti-atherogenic therapeutic approach, but it has been extremely challenging. Here, using in vivo phage display approach and the partial carotid ligation model of flow-induced atherosclerosis in mouse, we identified novel peptides that specifically bind to endothelial cells (ECs) exposed to disturbed flow condition in pro-atherogenic regions. https://www.leica-microsystems.com//science-lab/discovery-of-novel-peptides-targeting-pro-atherogenic-endothelium-in-disturbed-flow-regions-targeted-sirna-delivery-to-pro-atherogenic-endothelium-in-vivo/ Thu, 10 Nov 2016 15:28:00 +0000 https://www.leica-microsystems.com/18940 EM Sample Preparation Application Note for Leica EM ICE, Leica EM AFS2 - Electrical stimulation of neurons combined with high-pressure freezing allows physiological activation of synaptic activity and precise control over the time frame of the induced synaptic activity. https://www.leica-microsystems.com//science-lab/visualization-of-membrane-dynamics-with-millisecond-temporal-resolution/ Mon, 07 Nov 2016 10:53:00 +0000 PhD Shigeki Watanabe https://www.leica-microsystems.com/18213 EM Sample Preparation Application Note for Leica EM RES102 - The vertical layer construction of a semiconductor structure should be examined as a TEM cross-sectional sample. In addition to the specific preparation of the desired structure, the widely different sputter rates and atomic weights of the individual components represent the level of difficulty involved with this preparation problem. https://www.leica-microsystems.com//science-lab/cross-sectional-preparation-of-structured-semiconductor-materials-for-tem/ Mon, 31 Oct 2016 08:15:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18235 EM Sample Preparation Application Note for Leica EM RES102 - The surfaces of polished cross sections often show fine scratches and residues of the removed material or of the abrasive material. The artefacts are strongly material-dependent, and are mostly only detectable at higher resolutions in the scanning electron microscope. A further problem arises from the fact that the ground section mostly only has low contrast, i.e., in the structures of the semiconductor materials are very difficult to discern. With the use of ion beam milling, the ground sections of semiconductor structures can be "contrasted". https://www.leica-microsystems.com//science-lab/contrast-enhancement-of-polished-cross-sections-of-semiconductor-structures-sample-preparation-for-sem/ Fri, 28 Oct 2016 12:08:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/17540 Stereo Microscopy Live-Cell Imaging Fluorescence Microscopy Among the aquatic model organisms used in molecular and developmental biology the most prominent are the zebrafish (genus species: Danio rerio), medaka or japanese rice fish (genus species: Oryzias latipes), and african clawed frog (genus species: Xenopus laevis). This report gives useful information to scientists and technicians which can help improve their daily laboratory work by making the steps of transgenesis, fluorescent screening, and functional imaging more efficient. https://www.leica-microsystems.com//science-lab/work-more-efficiently-in-developmental-biology-with-stereo-microscopy-zebrafish-medaka-and-xenopus/ Thu, 27 Oct 2016 07:50:00 +0000 PhD James DeRose, Anastasia Felker, Dr. Elena Chiavacci, Gianluca D’Agati, Ph.D. Christian Mosimann, Ph.D. Jens Peter Gabriel, PhD Heinrich Bürgers, Ph.D. Adam Cliffe https://www.leica-microsystems.com/18806 Super-Resolution Do you know that most protists and bacteria lack in one feature that each of our body cell has? Our cells are touch and communicate with one another. They send and receive a variety of signals that coordinate their behavior to act together as a functional multicellular organism. Exploring the way of cellular communication and the ways how the cell surface interacts to organize tissues and body structures is of great interest. Kees Jalink and his team of scientists at the Netherlands Cancer Institute (NKI) in Amsterdam obtained new scientific insights into the molecular architecture of hemidesmosomes, cytoskeletal components, cell surface receptors and vesicular proteins with the help of Ground-State-Depletion (GSD)/ dSTORM microscopy. In this interview, Kees Jalink comments on their developments in imaging chambers, buffer conditions and image analysis to get the perfect super resolution image. https://www.leica-microsystems.com//science-lab/practical-guide-for-excellent-gsdim-super-resolution-images/ Wed, 26 Oct 2016 06:58:00 +0000 PhD Kees Jalink, PhD Tamara Straube, MSc Leila Nahidiazar, MSc Daniela Leyton Puig https://www.leica-microsystems.com/18142 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of wall cress (Arabidopsis thaliana) pod with subsequent gold coating and SEM analysis. https://www.leica-microsystems.com//science-lab/wall-cress-pod-protocol-critical-point-drying-of-arabidopsis-thaliana-for-sem/ Tue, 25 Oct 2016 08:00:00 +0000 Dr. Chen LiYu https://www.leica-microsystems.com/18919 Super-Resolution Protrusion of lamellipodia and ruffles requires polymerization of branched actin filaments by the Arp2/3 complex. Although regulation of Arp2/3 complex activity has been extensively investigated, the mechanism of initiation of lamellipodia and ruffles remains poorly understood. Here, we show that mDia1 acts in concert with the Arp2/3 complex to promote initiation of lamellipodia and ruffles. https://www.leica-microsystems.com//science-lab/initiation-of-lamellipodia-and-ruffles-involves-cooperation-between-mdia1-and-the-arp23-complex/ Mon, 24 Oct 2016 11:47:00 +0000 https://www.leica-microsystems.com/18915 Super-Resolution Elastin is an essential protein found in a variety of tissues where resilience and flexibility are needed, such as the skin and the heart. When aiming to engineer suitable implants, elastic fibres are needed to allow adequate tissue renewal. However, the visualization of human elastogenesis remains in the dark. To date, the visualization of human tropoelastin (TE) production in a human cell context and its fibre assembly under live cell conditions has not been achieved. Here, we present a long-term cell culture model of human dermal fibroblasts expressing fluorescence-labelled human TE. We employed a lentiviral system to stably overexpress Citrine-labelled TE to build a fluorescent fibre network. Using immunofluorescence, we confirmed the functionality of the Citrine-tagged TE. Furthermore, we visualized the fibre assembly over the course of several days using confocal microscopy. Applying super resolution microscopy, we were able to investigate the inner structure of the elastin–fibrillin-1 fibre network. https://www.leica-microsystems.com//science-lab/visualizing-tropoelastin-in-a-long-term-human-elastic-fibre-cell-culture-model/ Mon, 24 Oct 2016 10:10:00 +0000 https://www.leica-microsystems.com/18900 Super-Resolution Confocal Microscopy Application example of HvYolution Super-Resolution - Drug resistance, metastasis, and a mesenchymal transcriptional program are central features of aggressive breast tumors. The GTPase Arf6, often overexpressed in tumors, is critical to promote epithelial–mesenchymal transition and invasiveness. The metabolic mevalonate pathway (MVP) is associated with tumor invasiveness and known to prenylate proteins, but which prenylated proteins are critical for MVP-driven cancers is unknown. We show here that MVP requires the Arf6-dependent mesenchymal program. https://www.leica-microsystems.com//science-lab/p53-and-mevalonate-pathway-driven-malignancies-require-arf6-for-metastasis-and-drug-resistance/ Wed, 19 Oct 2016 16:27:00 +0000 https://www.leica-microsystems.com/18905 Super-Resolution Neuroscience Neuronal communication relies on synaptic vesicles undergoing regulated exocytosis and recycling for multiple rounds of fusion. Whether all synaptic vesicles have identical protein content has been challenged, suggesting that their recycling ability may differ greatly. Botulinum neurotoxin type-A (BoNT/A) is a highly potent neurotoxin that is internalized in synaptic vesicles at motor nerve terminals and induces flaccid paralysis. Recently, BoNT/A was also shown to undergo retrograde transport, suggesting it might enter a specific pool of synaptic vesicles with a retrograde trafficking fate. Using high-resolution microscopy techniques including electron microscopy and single molecule imaging, we found that the BoNT/A binding domain is internalized within a subset of vesicles that only partially co-localize with cholera toxin B-subunit and have markedly reduced VAMP2 immunoreactivity. https://www.leica-microsystems.com//science-lab/botulinum-neurotoxin-type-a-enters-a-non-recycling-pool-of-synaptic-vesicles/ Wed, 19 Oct 2016 11:51:00 +0000 https://www.leica-microsystems.com/18876 Super-Resolution The 6th European Super-Resolution User Club Meeting was held in collaboration with Dr. Timo Zimmermann, CRG, and Dr. Pablo Loza-Alvarez, ICFO, Barcelona. According to the founding principle of the club of keeping close to science, both imaging facilities at the CRG and the ICFO opened their doors to the User Club members, allowing them to explore exciting super-resolution and and nanoscopy applications. The meeting agenda covered highly relevant talks around this year’s central theme “Core Facilities and Super-Resolution Microscopy”, as well as plenty of opportunities to network amongst super-resolution users from different European countries. Here we present the abstracts of the talks held during the meeting. https://www.leica-microsystems.com//science-lab/abstracts-of-the-6th-european-super-resolution-user-club-meeting/ Tue, 18 Oct 2016 08:59:00 +0000 Dr. Timo Zimmermann, Dr. Pablo Loza-Alvarez, Dr. Alberto Lleó, Dr. Gražvydas Lukinavicius, Prof. Philip Tinnefeld, Prof. Hans-Georg Kräusslich, Dr. Steffen Dietzel, Dr. Valeria Caiolfa, Lorenzo Albertazzi https://www.leica-microsystems.com/18257 EM Sample Preparation Application Note for Leica EM RES102 - By means of cleaning, polishing and contrast enhancement a soft ZnAg sample should be prepared to obtain information concerning the grain structure and interfaces of the sample. The sample is contaminated after mechanical polishing. There are still some scratches on the surface. Grain structure is almost invisible. https://www.leica-microsystems.com//science-lab/surface-modification-of-znag-sample-sample-preparation-for-sem/ Mon, 17 Oct 2016 08:16:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18364 EM Sample Preparation Application Note for Leica EM TIC 3X - Electron Backscattered Diffraction (EBSD) is a surface technique creating a diffraction pattern (Kikuchi-bands). It can be used for crystal orientation mapping, defect studies, phase identification, grain boundary studies and morphology studies. The information depth is just a few nm. Therefore a good sample preparation is very important. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-nicu-on-steel-for-ebsd/ Fri, 14 Oct 2016 09:45:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18096 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of E. coli with subsequent platinum / palladium coating and SEM analysis. Sample was inserted into a filter disc (Pore size: 16 - 40 μm) and placed into the filter discs and porous pots holder. Cultivate fungi and bacteria on agar containing growth medium for 3 days. https://www.leica-microsystems.com//science-lab/bacteria-protocol-critical-point-drying-of-e-coli-for-sem/ Thu, 13 Oct 2016 10:04:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/18881 Surgical Microscopy In endodontics, accurate treatment is not only dependent on the technical skills and knowledge of the dentist, but also on clear, detailed visualization of the surgical field. As the outcome of an endodontic therapy is influenced by many factors that are not visible to the naked eye – e.g. additional root canals or anatomical variations – the high magnification and illumination provided by a dental microscope has become indispensable for both diagnosis and therapy. Today, it is widely agreed that the use of dental microscopes has helped to extend endodontic treatment potential. https://www.leica-microsystems.com//science-lab/successful-endodontic-treatment-supported-by-a-dental-microscope/ Tue, 11 Oct 2016 08:30:00 +0000 https://www.leica-microsystems.com/18178 EM Sample Preparation Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve flat cuts of material combinations consisting of hard and soft materials. Here, cross sectioning of painted concrete is describes in order to visualize the interface between paint and concrete. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-painted-concrete/ Mon, 10 Oct 2016 20:39:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18104 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of nuclear pores from clawed frog oocytes with subsequent chromium coating and SEM analysis. Silicon chips containing the samples were placed into the filter discs and porous pots holder. https://www.leica-microsystems.com//science-lab/clawed-frog-nuclear-envelope-protocol/ Thu, 06 Oct 2016 16:00:00 +0000 Dr. Martin W. Goldberg, M.Sc. Christine Richardson https://www.leica-microsystems.com/18092 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of the blow fly with subsequent X-ray micro-computed tomography (micro-CT) to detect neuroanatomical features. https://www.leica-microsystems.com//science-lab/micro-computed-tomography-micro-ct-of-insect-brain-protocol/ Thu, 06 Oct 2016 14:41:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/18259 EM Sample Preparation Application Note for Leica EM RES102 - Thin foils are mostly unstable because of their thickness of a few microns. This makes it difficult to do slope cutting without any protection of the sample. A common realisation to protect the sample surface is by sticking a cover glass on top of the sample. Another issue is cutting the foils before ion milling. The sample edge should be flat and sharp without any broken areas. A razor blade is mostly the best solution. A protected sample can salso be sawed with a wire saw. https://www.leica-microsystems.com//science-lab/thin-metal-foils-with-coatings-sample-preparation-for-sem/ Wed, 05 Oct 2016 07:16:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/17476 Stereo Microscopy Live-Cell Imaging Fluorescence Microscopy For scientists and technicians working with fruit flies, most often genus Drosophila, this report is intended to give useful information to help improve daily laboratory work by making the steps of fly pushing, fluorescent screening, dissection, and documentation/imaging more efficient. It also details various possibilities for properly equipping or stocking a fly lab. https://www.leica-microsystems.com//science-lab/work-more-efficiently-in-developmental-biology-with-stereo-microscopy-fruit-flies-drosophila-melanogaster/ Thu, 22 Sep 2016 08:58:00 +0000 PhD Heinrich Bürgers, PhD James DeRose, Ph.D. Adam Cliffe https://www.leica-microsystems.com/18818 EM Sample Preparation Application Note for Leica EM ACE600 - Recent developments in aberration corrected transmission electron microscopes as well as further improvements in monochromaters and spectrometers have pushed the attainable energy resolution for Electron energy loss spectroscopy (EELS) to 100 meV and beyond. STEM-EELS of individual nanomaterials can be challenging due the necessity of a support film. https://www.leica-microsystems.com//science-lab/ultra-thin-carbon-support-films-for-improved-stem-eels-analysis-of-nanoparticles/ Wed, 21 Sep 2016 11:27:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/18820 EM Sample Preparation Application Note for Leica EM ACE600 - Much of the battle involved in obtaining good transmission electron microscopy data is in the specimen preparation itself. Even though some nanomaterials are already electron transparent (e.g. nanoparticles and proteins) and often do not require further thinning procedures, they need to be dispersed onto thin support films. https://www.leica-microsystems.com//science-lab/ways-to-reveal-more-from-your-samples-ultra-thin-carbon-films/ Wed, 21 Sep 2016 11:23:00 +0000 PhD Frédéric Leroux, Jan de Weert https://www.leica-microsystems.com/18801 Super-Resolution Fluorescence Microscopy A new type of fluorescent bead is presented by GATTAquant. These beads, called GATTA-Beads, are characterized by a small diameter (23 nm), high intensity and size uniformity. In combination with state-of the-art STED microscopes such as the Leica TCS SP8 STED 3X and high-end image restoration methods available in the Huygens Software, it is shown that these new beads can be used for accurate STED PSF characterization in 3D. Furthermore, it is shown that the measured 3D STED-PSF can be used to improve image restoration quality in combination with STED deconvolution methods available in the Huygens Software. https://www.leica-microsystems.com//science-lab/measuring-the-3d-sted-psf-with-a-new-type-of-fluorescent-beads/ Wed, 21 Sep 2016 06:54:00 +0000 PhD Jürgen J. Schmied, MSc Remko Dijkstra, Ph.D. Max B. Scheible, Ph.D. Giulia M. R. De Luca, PhD Jochen J. Sieber https://www.leica-microsystems.com/18192 EM Sample Preparation Application Note for Leica EM TIC 3X - Purpose: The shape of the wire is difficult for ion beam slope cutting. Goal: Cross sectional preparation to see the structure of the wire. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-a-superconductive-wire/ Tue, 20 Sep 2016 16:41:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18691 Stereo Microscopy Confocal Microscopy Fluorescence Microscopy For scientists, technicians, and teachers working with the worm C. elegans in the research lab or classroom, this report is intended to give useful information to help improve their daly work. The aim is to make the work steps of worm picking, transgenesis, RNA interference, screening, and functional imaging efficient. It also details the various possibilities for equipping a research worm lab or biology classroom/teaching lab explaining worm methods. https://www.leica-microsystems.com//science-lab/work-efficiently-in-developmental-biology-with-stereo-and-confocal-microscopy-c-elegans/ Mon, 19 Sep 2016 06:09:00 +0000 PhD James DeRose, PhD Heinrich Bürgers, PhD Martin Gamerdinger https://www.leica-microsystems.com/18149 EM Sample Preparation Application Note for Leica EM ICE, Leica EM AFS2 - Wildtype L4 stage C. elegans (N2 strain) were placed in the 100 μm deep side of Lecithin-coated (see detailed protocol*) type A 3 mm Cu/Au carriers (Leica) with extracellular filler containing 1% (w/v) Agarose type IX and 2% (w/v) Bovine Serum Albumin in bacteria medium (see preparation details**) and sandwiched with the flat side of Lecithin-coated type B 3 mm Cu/Au carriers (Leica). Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/structural-study-of-c-elegans/ Fri, 16 Sep 2016 08:15:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. Rob Mesman https://www.leica-microsystems.com/18344 EM Sample Preparation Application Note for Leica EM TIC020, Leica EM TIC 3X - Electron backscattered diffraction (EBSD) is for example used to examine the crystallographic orientation of material. The sample preparation for such samples is sometimes very tricky as the depth of information is just few nm (~20nm or less). That means the sample surface must be flat and free of preparation artefacts. Mechanical polishing leads mostly to sample surfaces damages. https://www.leica-microsystems.com//science-lab/cross-section-of-an-aluminium-sample-for-electron-backscattered-diffraction-ebsd/ Thu, 15 Sep 2016 07:54:00 +0000 Robert Ranner https://www.leica-microsystems.com/18779 Multiphoton Microscopy Multipohoton microscopy experiment using Leica TCS SP8 MP and Leica 20x/0.95 NA BABB immersion objective. Understanding kidney microanatomy is key to detecting and identifying early events in kidney disease. Improvements in tissue clearing and imaging have been crucial in this field, and now we report on a novel, time-efficient method to study podocyte depletion in renal glomeruli using a combination of immunofluorescence, optical clearing, confocal microscopy and 3D analysis. https://www.leica-microsystems.com//science-lab/babb-clearing-and-imaging-for-high-resolution-confocal-microscopy-counting-and-sizing-kidney-cells-in-the-21st-century/ Tue, 13 Sep 2016 09:03:00 +0000 Ph.D. Victor G. Puelles, Prof. John F. Bertram, MSc Stephen Firth, Prof. Ian Harper https://www.leica-microsystems.com/18796 Multiphoton Microscopy Confocal Microscopy Light Sheet Microscopy Neuroscience Chemical clearing of fixed tissues is becoming a key instrument for the three-dimensional reconstruction of macroscopic tissue portions, including entire organs. Indeed, the growing interest in this field has both triggered and been stimulated by recent advances in high-throughput microscopy and data analysis methods, which allowed imaging and management of large samples. https://www.leica-microsystems.com//science-lab/clearing-of-fixed-tissue-a-review-from-a-microscopists-perspective/ Mon, 12 Sep 2016 11:20:00 +0000 https://www.leica-microsystems.com/18301 EM Sample Preparation Application Note for Leica EM HPM100 - Frozen samples (90 μm thick slices frozen by HPM100) were inserted into a double replica table and then fractured into two pieces at –130°C (after insertion of the tissue into BAF 060 the samples should be left in the chamber for 20 min to reach the –130°C). https://www.leica-microsystems.com//science-lab/freeze-fracture-replication-of-pyramidal-cells/ Thu, 08 Sep 2016 16:23:00 +0000 Akos Kulik https://www.leica-microsystems.com/18112 Confocal Microscopy The congenital malformation split hand/foot (SHFM) is characterized by missing central fingers and dysmorphology or fusion of the remaining ones. During limb development, the apical ectodermal ridge (AER) is a key-signaling center responsible for early proximal–distal growth and patterning. In search for the mechanism, we examined the non-canonical Wnt signaling, considering that Dwnt-5 is a target of distalless in Drosophila and the knockout of Wnt5, Ryk, Ror2 and Vangl2 in the mouse causes severe limb malformations. https://www.leica-microsystems.com//science-lab/the-apical-ectodermal-ridge-of-the-mouse-model-of-ectrodactyly-dlx5dlx6-shows-altered-stratification-and-cell-polarity-which-are-restored-by-exogenous-wnt5a-ligand/ Wed, 07 Sep 2016 07:18:00 +0000 https://www.leica-microsystems.com/18239 EM Sample Preparation Application Note for Leica EM RES102 - Ion milling can be used to reduce the roughness of sample surfaces. Small angles less than 6° with respect to the sample surface are necessary. The high voltage depends on the material to be prepared. The reason for the levelling effect is the different milling angle of flat and rough surface areas. The milling rate is lower for small angles. The rough surface area will be faster milled. Ion polishing is often the final step of sample preparation. The prerequisite is a perfect mechanical prepreparation as samples with deep surface scratches cannot be ion polished. Soft materials usually have a smeared sample surface after mechanical polishing. It is necessary to remove this smeared material before ion polishing. Otherwise the above mentioned polishing effect does not work. https://www.leica-microsystems.com//science-lab/ion-beam-polishing-of-sample-surfaces-sample-preparation-for-sem/ Tue, 06 Sep 2016 11:11:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18730 Confocal Microscopy Live-Cell Imaging Adeno-associated viral (AAV) vectors are becoming an important tool for gene therapy of numerous genetic and other disorders. Several recombinant AAV vectors (rAAV) have the ability to transduce striated muscles in a variety of animals following intramuscular and intravascular administration, and have attracted widespread interest for therapy of muscle disorders such as the muscular dystrophies. Here we examined the relative ability of rAAV vectors derived from AAV6 to target myoblasts, myocytes, and myotubes in culture and satellite cells and myofibers in vivo. AAV vectors are able to transduce proliferating myoblasts in culture, albeit with reduced efficiency relative to postmitotic myocytes and myotubes. In contrast, quiescent satellite cells are refractory to transduction in adult mice. https://www.leica-microsystems.com//science-lab/adeno-associated-viral-vectors-do-not-efficiently-target-muscle-satellite-cells/ Mon, 05 Sep 2016 05:58:00 +0000 https://www.leica-microsystems.com/18215 EM Sample Preparation Application Note for Leica EM RES102 - Mechanical polishing of soft materials or hard / soft material combinations is tricky. The mechanical polishing process leads very often to smearing of the soft material. The smeared material covers the surface and fills small pores or holes. Grain structures, interfaces and other structural details can be masked. An additional ion milling step with milling angles between 10° C and 15° C with respect to the sample surface can remove or reduce the contamination. https://www.leica-microsystems.com//science-lab/cleaning-of-smeared-sample-surfaces-sample-preparation-for-sem/ Fri, 02 Sep 2016 06:49:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18090 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of book scorpion with subsequent X-ray micro-computed tomography (micro-CT) to detect anatomical features with special regard to the musculature. https://www.leica-microsystems.com//science-lab/micro-computed-tomography-micro-ct-of-book-scorpion-musculature/ Thu, 01 Sep 2016 08:39:00 +0000 PhD Elisabeth Lipke, PhD Peter Michalik https://www.leica-microsystems.com/18146 EM Sample Preparation Application Note for Leica EM ICE - Rat Hippocampal neurons, cultured on 50 μm thick Aclar (Aclar embedding film, EMS) for 19 days, were frozen in the 100 μm deep side of lecithin coated (detailed protocol Appendix I) type A 3 mm Cu/Au carriers (Leica) and sandwiched with the flat side of lecithin coated type B 3 mm Cu/Au carriers (Leica). No additional filler was used, only cell culture medium with the addition of Hepes buffer pH 7.2 to a final concentration of 25 mM. Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/cultured-rat-hippocampal-neurons/ Mon, 29 Aug 2016 07:12:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. W. H. Mueller, Prof. Dr. C. C. Hoogenraad https://www.leica-microsystems.com/18742 Basics in Microscopy Widefield Microscopy The number of applications in microscopy can be as manifold as life itself. Different specimens demand different contrast methods to get a decent result. Unstained cells normally do not show a very high contrast. Researchers approach this problem by either staining the specimen with dyes respectively fluorescent molecules; or adding contrast by manipulating the light passing through. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-application-options-with-leica-dm2500-led-microscope/ Tue, 23 Aug 2016 11:45:00 +0000 Dr. Christoph Greb, PhD Tamara Straube https://www.leica-microsystems.com/18000 EM Sample Preparation Application Note for Leica EM AMW - The tissues were fixed in the modified Karnovsky fixative generally by immersion overnight (at minimum 4h at room temperature). Then pieces of approx. 1mm3 were cut with a sharp razor blade and processed for embedding in the Leica EM AMW Microwave Tissue Processor. https://www.leica-microsystems.com//science-lab/epoxy-resin-embedding-of-animal-and-human-tissues-for-pathological-diagnosis-and-research/ Fri, 19 Aug 2016 08:34:00 +0000 Dr. Josef A. Schroeder https://www.leica-microsystems.com/19026 Super-Resolution Axial excitation confinement beyond the diffraction limit is crucial to the development of next-generation, super-resolution microscopy. STimulated Emission Depletion (STED) nanoscopy offers lateral super-resolution using a donut-beam depletion, but its axial resolution is still over 500 nm. Total internal reflection fluorescence microscopy is widely used for single-molecule localization, but its ability to detect molecules is limited to within the evanescent field of ~100 nm from the cell attachment surface. We find here that the axial thickness of the point spread function (PSF) during confocal excitation can be easily improved to 110 nm by replacing the microscopy slide with a mirror. The interference of the local electromagnetic field confined the confocal PSF to a 110-nm spot axially, which enables axial super-resolution with all laser-scanning microscopes. https://www.leica-microsystems.com//science-lab/mirror-enhanced-super-resolution-microscopy/ Thu, 18 Aug 2016 09:29:00 +0000 https://www.leica-microsystems.com/13942 Fluorescence Microscopy Quantitative Imaging In this talk, Roger Tsien discusses how fluorescent proteins have been turned into indicators for a wide variety of biological molecules, including pH, ions, redox potential, and signaling molecules like phosphoinositides. The talk also covers reporters used to measure the activity of enzymes like kinases, phosphatases, and proteases. It covers both single proteins whose intensity or wavelength change, as well as reporters using Förster resonance energy transfer (FRET). https://www.leica-microsystems.com//science-lab/video-talk-by-roger-tsien-fluorescent-protein-indicators/ Tue, 16 Aug 2016 10:29:00 +0000 PhD Roger Y. Tsien https://www.leica-microsystems.com/18209 EM Sample Preparation Application Note for Leica EM RES102 - Ceramic samples are mostly very brittle, and are therefore very difficult to thin mechanically to a low starting thickness for ion beam milling. The ion beam milling of insulators often leads to static charging of the surface of the sample. This, in turn, reduces the sputter rate. When using the Ti standard holder (standard TEM holder), however, sufficient secondary electrons are created by the ion beam also falling on the sample holder to largely compensate for the static charging. https://www.leica-microsystems.com//science-lab/ceramics-sample-preparation-for-tem/ Mon, 15 Aug 2016 18:46:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18295 EM Sample Preparation Application Note for Leica EM HPM100 - The sections are of yeast frozen with a Leica HPM100 high pressure freezer in the copper tube system, the cell paste was mixed with a pH 6.5 MES/dextran buffer so that a final MES concentration of 50mM and a dextran concentration of 20% was achieved. https://www.leica-microsystems.com//science-lab/cryo-electron-microscopy-of-vitreous-sections-cemovis-of-yeast/ Fri, 12 Aug 2016 14:51:00 +0000 Dr. Jonathan O'Driscoll, Dr. Daniel Kofi Clare, Prof. Helen Saibil https://www.leica-microsystems.com/18144 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of wrinkled giant hyssop leaf with subsequent gold coating and SEM analysis. https://www.leica-microsystems.com//science-lab/wrinkled-giant-hyssop-leaf-protocol/ Thu, 11 Aug 2016 17:04:00 +0000 Dr. Guo JianSheng https://www.leica-microsystems.com/18102 Super-Resolution The ability of invariant natural killer T (iNKT) cells to recognize endogenous antigens represents a distinct immune recognition strategy, which underscores the constitutive memory phenotype of iNKT cells and their activation during inflammatory conditions. By using superresolution microscopy, we show that CD1d molecules form nanoclusters at the cell surface of APCs, and their size and density are constrained by the actin cytoskeleton. https://www.leica-microsystems.com//science-lab/the-actin-cytoskeleton-modulates-the-activation-of-inkt-cells-by-segregating-cd1d-nanoclusters-on-antigen-presenting-cells/ Wed, 10 Aug 2016 13:28:00 +0000 https://www.leica-microsystems.com/18100 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of Black mould (Aspergilus niger) with subsequent platinum / palladium coating and SEM analysis to detect conidiospores. Sample was inserted into a filter disc (Pore size: 16 - 40 μm) and placed into the filter discs and porous pots holder. https://www.leica-microsystems.com//science-lab/black-mould-protocol/ Tue, 09 Aug 2016 12:01:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/18151 EM Sample Preparation Application Note for Leica EM TIC 3X - Alumina is very difficult to handle and almost impossible to prepare with conventional methods. Ion beam slope cutting is a method that can achieve cross sections of material combinations consisting of hard and soft components. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-alumina/ Mon, 08 Aug 2016 07:48:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18115 Confocal Microscopy A human gut-on-a-chip microdevice was used to coculture multiple commensal microbes in contact with living human intestinal epithelial cells for more than a week in vitro and to analyze how gut microbiome, inflammatory cells, and peristalsis-associated mechanical deformations independently contribute to intestinal bacterial overgrowth and inflammation. This in vitro model replicated results from past animal and human studies, including demonstration that probiotic and antibiotic therapies can suppress villus injury induced by pathogenic bacteria. https://www.leica-microsystems.com//science-lab/contributions-of-microbiome-and-mechanical-deformation-to-intestinal-bacterial-overgrowth-and-inflammation-in-a-human-gut-on-a-chip/ Fri, 05 Aug 2016 16:29:00 +0000 https://www.leica-microsystems.com/17998 EM Sample Preparation Application Note for Leica EM AMW - Plants were grown in growth chambers under defined conditions. After stratification for 4 days at 4°C seeds were grown in pots with soil with 9/15 hours day/night photoperiod. Day and night temperatures were 22°C and 18°C, respectively, the relative humidity was 60% and the plants were kept at 100% relative soil water content. Light intensity varied between 110 and 140 μmol m-2 s-1. https://www.leica-microsystems.com//science-lab/arabidopsis-thalianal-accession-col/ Mon, 01 Aug 2016 13:03:00 +0000 Prof. Bernd Zechmann, Prof. Günther Zellnig https://www.leica-microsystems.com/18190 EM Sample Preparation Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve flat cross sections of soft materials or material combinations consisting of hard and soft components. The CuSn connector is very soft. Mechanical polishing leads to smearing. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-cusn-connector-of-a-solar-cell/ Fri, 29 Jul 2016 11:56:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/15428 Confocal Microscopy Live-Cell Imaging H2S is the third endogenously generated gaseous signaling compound and has also been known to involve a variety of physiological processes. To better understand its physiological and pathological functions, efficient methods for monitoring of H2S are desired. Azide fluorogenic probes are popular because they can take place bioorthogonal reactions. In this work, by employing a fluorescein derivative as the fluorophore and an azide group as the recognition unit, we reported a new probe 5-azidofluorescein for H2S with improved sensitivity and selectivety. https://www.leica-microsystems.com//science-lab/highly-selective-fluorescent-and-colorimetric-probe-for-live-cell-monitoring-of-sulphide-based-on-bioorthogonal-reaction/ Fri, 29 Jul 2016 11:41:00 +0000 https://www.leica-microsystems.com/17987 EM Sample Preparation Application Note for Leica EM AFS2 - Mouse heart tissue from wild-type (WT) and αT -catenin KO (KO) mice was excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer Leica EM PACT. Freeze subs­titution was carried out using a Leica EM AFS in dry acetone containing 2% ddH2O and 0.1% glutaraldehyde over a 4-days period. https://www.leica-microsystems.com//science-lab/immuno-electron-microscopy-of-high-pressure-frozen-and-freeze-substituted-mouse-heart/ Wed, 27 Jul 2016 15:35:00 +0000 Riet de Rycke https://www.leica-microsystems.com/17979 EM Sample Preparation Application Note for Leica EM AFS2 - Cells of the the moderately thermophilic Acidithiobacillus sp.strain HV2/2, were centrifuged at 20,0009xg and then cryo-immobilized by high-pressure freezing on gold carriers. https://www.leica-microsystems.com//science-lab/cell-envelope-structure-of-a-gram-negative-thermotolerant-y-proteobacterium-acidithiobacillus-sp-strain-hv22-and-its-interaction-with-pyrite/ Mon, 25 Jul 2016 10:12:00 +0000 Dr. Andreas Klingl https://www.leica-microsystems.com/18088 Super-Resolution Super-resolution microscopy is transforming our understanding of biology but accessibility is limited by its technical complexity, high costs and the requirement for bespoke sample preparation. We present a novel, simple and multi-color super-resolution microscopy technique, called translation microscopy (TRAM), in which a super-resolution image is restored from multiple diffraction-limited resolution observations using a conventional microscope whilst translating the sample in the image plane. https://www.leica-microsystems.com//science-lab/translation-microscopy-tram-for-super-resolution-imaging/ Fri, 22 Jul 2016 17:51:00 +0000 https://www.leica-microsystems.com/18049 EM Sample Preparation Neuroscience Application Note for Leica EM ICE - WT hippocampal neurons were plated at a density of 80,000 cells/cm2 on 6 mm sapphire disks for 14 days. Sample were frozen using a high-pressure freezer (Leica EM ICE) under a pressure of 2100bar by mounting it into a sandwich support with extracellular solution containing 15% of Ficoll 400, to assess ice crystal damage. The Cryo-fixation was achieved within milliseconds allowing simultaneous immobilization of all macromolecular components. After freezing, sam­ple was transferred into cryovials containing 1% glutaraldehyde, 1% osmium tetroxide, 1% milliQwater in anhydrous acetone and processed in an automated freeze-substitution device (Leica EM AFS2). https://www.leica-microsystems.com//science-lab/symmetric-synapse-clathrin-coated-endocytosis-pit-in-the-postsynaptic-dendrite/ Wed, 20 Jul 2016 08:56:00 +0000 Dr. Shuwen Chang https://www.leica-microsystems.com/18623 Fluorescence Microscopy CRISPR-Cas9 The CRISPR-Cas system was originally discovered as an adaptive immune system of bacteria and archaea to protect against viral attack. During this talk, given at Leica Microsystems in Wetzlar, Dr. Lennart Randau, MPI Marburg, presents this simple and fascinating system in detail. Furthermore, he introduces the adaption of the CRISPR-Cas system into a potent molecular biology tool, which is used heavily for genome editing. In addition to its influence on molecular biology, meanwhile the Cas9 nuclease also stimulates microscopy techniques e.g. to fluorescently label genomic loci in living cells. https://www.leica-microsystems.com//science-lab/video-talk-crispr-cas-from-a-bacterial-adaptive-immune-system-to-a-genome-engineering-tool/ Tue, 12 Jul 2016 04:13:00 +0000 Dr. Lennart Randau https://www.leica-microsystems.com/18271 Super-Resolution Quantitative Imaging Live-Cell Imaging Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as of lipids in plasma membrane bioactivity. However, these dynamics can often only be visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, an extension of fluorescence correlation spectroscopy, FCS) on a super-resolution stimulated emission depletion (STED) microscope, we here extend previous observations of nanoscale lipid dynamics in the plasma membrane of living mammalian cells. https://www.leica-microsystems.com//science-lab/sted-flcs-an-advanced-tool-to-reveal-spatiotemporal-heterogeneity-of-molecular-membrane-dynamics/ Mon, 11 Jul 2016 08:53:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/17975 EM Sample Preparation Application Note for Leica EM AFS2 - Arabidopsis thaliana roots (mutant PIN1pro:PIN1-GFP;bex5-1) were excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer. Freeze substitution was carried out using a Leica EM AFS2. https://www.leica-microsystems.com//science-lab/electron-microscopy-of-high-pressure-frozen-and-freeze-substituted-arabidopsis-thaliana-root-tips-cells/ Fri, 08 Jul 2016 10:02:00 +0000 Riet de Rycke https://www.leica-microsystems.com/15645 Confocal Microscopy Live-Cell Imaging We report a new technique for high-resolution in vivo imaging of myelinated axons in the brain, spinal cord and peripheral nerve that requires no fluorescent labeling. This method, based on spectral confocal reflectance microscopy (SCoRe), uses a conventional laser scanning confocal system to generate images by merging the simultaneously reflected signals from multiple lasers of different wavelengths. https://www.leica-microsystems.com//science-lab/label-free-in-vivo-imaging-of-myelinated-axons-in-health-and-disease-with-spectral-confocal-reflectance-microscopy/ Fri, 01 Jul 2016 09:29:00 +0000 https://www.leica-microsystems.com/18368 EM Sample Preparation Application Note for Leica TIC 3X - High quality sample preparation of large area to investigate the sample in the SEM. For the mechanical preparation step diamond lapping foils of 9μm subsequently 2μm and finally 0.5μm grain size were used. It took about 1.5 hours. The sample was removed from the stub with a razor blade after TXP processing and fixed onto the holder of the rotary stage of the Leica EM TIC 3X. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-oil-shale-rock/ Tue, 28 Jun 2016 13:04:00 +0000 Robert Ranner https://www.leica-microsystems.com/18431 Quantitative Imaging Live-Cell Imaging There is solid evidence for analyzing fluorescence correlation and dual color fluorescence crosscorrelation spectroscopy data (FCS and dual color FCCS) in cellular applications by equations based on anomalous subdiffusion. Using equations based on normal diffusion causes artifacts of the fitted biological system response parameters and of the interpretations of the FCS and dual color FCCS data in the crowded environment of living cells. Equations based on normal diffusion are not valid in living cells. The original article embraces the status of the experimental situation and touches obstacles that still hinder the applications of single molecules in the cellular environment. https://www.leica-microsystems.com//science-lab/individual-macromolecule-motion-in-a-crowded-living-cell/ Mon, 27 Jun 2016 04:57:00 +0000 https://www.leica-microsystems.com/18307 Surgical Microscopy Excellent red reflex as well as maximum accuracy throughout the entire procedure are required to meet these expectations. Dr. Ulrich Jung, Medical Director at the ARTEMIS eye clinic in Dillenburg, Germany, tested the Proveo 8 ophthalmological microscope from Leica Microsystems in combination with the positioning system IOLcompass Pro for intraocular lenses at his clinic. In this interview, Dr. Jung reports his experiences. https://www.leica-microsystems.com//science-lab/the-excellent-red-reflex-combined-with-the-iol-guidance-system-makes-it-possible-for-me-to-work-quickly-and-precisely-during-refractive-procedures/ Fri, 24 Jun 2016 11:40:00 +0000 Dr. med. Ulrich Jung, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/18309 Surgical Microscopy Successful refractive surgery is all about accuracy. The aim is to deliver minimal residual astigmatism so the patient can be glasses- free. Latest IOL Positioning Systems such as IOLcompass Pro deliver precise, dependable and dynamically optimized guidance information designed to minimize inaccuracy throughout the entire process. This comparison highlights the impact on a typical cataract surgery using toric IOLs. https://www.leica-microsystems.com//science-lab/how-to-manage-critical-points-in-cataract-or-refractive-surgery-with-iol-positioning-software/ Thu, 23 Jun 2016 16:44:00 +0000 https://www.leica-microsystems.com/17991 Super-Resolution We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave. https://www.leica-microsystems.com//science-lab/two-photon-excitation-sted-microscopy-with-time-gated-detection/ Tue, 21 Jun 2016 06:39:00 +0000 https://www.leica-microsystems.com/18108 Fluorescence Microscopy Confocal Microscopy The tumor-suppressive let-7 microRNA family targets various oncogene-encoding mRNAs. We identify the let-7 targets HMGA2, LIN28B and IGF2BP1 to form a let-7 antagonizing self-promoting oncogenic triangle. Surprisingly, 3′-end processing of IGF2BP1 mRNAs is unaltered in aggressive cancers and tumor-derived cells although IGF2BP1 synthesis was proposed to escape let-7 attack by APA-dependent (alternative polyadenylation) 3′ UTR shortening. https://www.leica-microsystems.com//science-lab/the-oncogenic-triangle-of-hmga2-lin28b-and-igf2bp1-antagonizes-tumor-suppressive-actions-of-the-let-7-family/ Fri, 17 Jun 2016 13:49:00 +0000 https://www.leica-microsystems.com/18284 EM Sample Preparation Application Note for Leica EM VCT100, Leica EM ACE600 - A thin layer of latex paint was spread on a clean, scored, silicon chip. The sample was immediately plunge frozen in liquid ethane and transferred under LN2 to the Leica EM VCT100 loading station where it was placed in the customized sample holder. https://www.leica-microsystems.com//science-lab/cryo-sem-imaging-of-latex-paint/ Thu, 16 Jun 2016 16:50:00 +0000 Dr. Levi Felts, Dr. Kim Rensing, Chris Frethem https://www.leica-microsystems.com/18184 EM Sample Preparation Application Note for Leica EM TIC 3X - Ion beam slope cutting is a method that can achieve cross sections of soft materials or material combinations consisting of hard and soft components. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-rubber-tire/ Tue, 14 Jun 2016 10:30:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/18164 Super-Resolution Collagen is the most abundant protein in the animal kingdom. It is of fundamental importance during development for cell differentiation and tissue morphogenesis as well as in pathological processes such as fibrosis and cancer cell migration. However, our understanding of the mechanisms of procollagen secretion remains limited. Here, we show that TFG organizes transitional ER (tER) and ER exit sites (ERESs) into larger structures. Depletion of TFG results in dispersion of tER elements that remain associated with individual ER-Golgi intermediate compartments (ERGICs) as largely functional ERESs. We show that TFG is not required for the transport and packaging of small soluble cargoes but is necessary for the export of procollagen from the ER. Our work therefore suggests a key relationship between the structure and function of ERESs and a central role for TFG in optimizing COPII assembly for procollagen export. https://www.leica-microsystems.com//science-lab/tfg-promotes-organization-of-transitional-er-and-efficient-collagen-secretion/ Tue, 14 Jun 2016 08:36:00 +0000 https://www.leica-microsystems.com/18251 Surgical Microscopy In dentistry a bright, magnified view into deep cavities supports detailed diagnosis and precise therapy, particularly in the field of endodontics. In this interview Dr. Dean Raicov explains the benefits of the dental microscope in his practice. Dr. Raicov is a dentist based in Röhrnbach, Germany and specializes in endodontics, parodontology, and prosthetics. He is a member of the European Society of Microscope Dentistry (ESMD) and the German Society of Endodontics and Paradontology. https://www.leica-microsystems.com//science-lab/the-dental-microscope-in-endodontics/ Mon, 13 Jun 2016 13:07:00 +0000 Dr. Dean Raicov https://www.leica-microsystems.com/18211 Surgical Microscopy These videos recorded during cataract and glaucoma surgery using Proveo 8 demonstrate how Dr. Ahmed benefits from continuous red reflex even during phaco, due to exclusive CoAx 4 coaxial illumination, and high resolution combined with depth of focus for a texture-rich view due to FusionOptics technology. https://www.leica-microsystems.com//science-lab/cataract-and-glaucoma-surgery-with-the-proveo-8-ophthalmic-microscope/ Wed, 08 Jun 2016 12:09:00 +0000 Dr. Ike Ahmed https://www.leica-microsystems.com/18086 EM Sample Preparation Application Note for Leica EM CPD300 - Critical point drying of water flea with subsequent gold coating and SEM-Analysis to detect finde surface structures. https://www.leica-microsystems.com//science-lab/water-flea-protocol/ Tue, 07 Jun 2016 07:06:00 +0000 Mag. Dr. Daniela Gruber https://www.leica-microsystems.com/17941 Laser Microdissection During Leica workshops for LMD users in Brazil, hosted by the Federal University of Paraná/UFPR (UFPR) at the Centro de Energia Nuclear na Agricultura/USP (CENA), the power of laser microdissection using the Leica LMD systems was demonstrated. One special focus was on plant dissection which needs a high laser power. https://www.leica-microsystems.com//science-lab/workflows-protocols-plant-laser-microdissection/ Mon, 06 Jun 2016 05:22:00 +0000 Dr. Falk Schlaudraff, Dr. Christoph Greb https://www.leica-microsystems.com/18077 Optical Coherence Tomography Surgical Microscopy Intraoperative ocular coherence tomography (iOCT) is a useful asset when performing macular surgery and other vitreoretinal procedures. Considering the advantages it provides, iOCT may be the next paradigm shift in vitreoretinal surgery according to Justis P. Ehlers, MD. https://www.leica-microsystems.com//science-lab/is-ioct-the-next-paradigm-shift-for-vitreoretinal-surgery/ Thu, 02 Jun 2016 14:19:00 +0000 https://www.leica-microsystems.com/18176 Confocal Microscopy Diabetes is quickly becoming the most widespread disorder in the Western world. Among the most prevalent effects of diabetes is atherosclerosis, which in turn is driven in part by inflammation. Both insulin and Tumor Necrosis Factor-alpha (TNFα) increase the presence of Vascular Cellular Adhesion Molecule-1 (VCAM-1) expression. The aim of this study is to determine the effects of downregulating Extracellular signal-Regulated Kinase-2 (ERK2) and Akt on insulin and TNFa-stimulated VCAM-1 expression. https://www.leica-microsystems.com//science-lab/erk2-and-akt-are-negative-regulators-of-insulin-and-tumor-necrosis-factor-a-stimulated-vcam-1-expression-in-rat-aorta-endothelial-cells/ Wed, 01 Jun 2016 12:14:00 +0000 https://www.leica-microsystems.com/18124 Quality Assurance EM Sample Preparation Today’s automotive industry use a variety of decorative and functional treatment to improve the vehicles surfaces. Traditional quality control methods to inspect these multilayer samples have proven to be extremely time-consuming and bear the risk of missing defects. A new approach combining a target surface system and a light microscope offers new possibilities of speed and reliability. F. Javier Ruiz Balbas, Laboratory Manager at Atotech Spain, explains his experiences with the system. https://www.leica-microsystems.com//science-lab/fjavier-ruiz-balbas-atotec-the-leica-system-offers-flexibility-and-permissively-to-achieve-high-quality-results-in-short-time/ Mon, 30 May 2016 13:00:00 +0000 F.Javier Ruiz Balbas https://www.leica-microsystems.com/18079 EM Sample Preparation Application Note for Leica EM CPD300 - Life Science Research. Species: Human (Homo sapiens) Critical point drying of human blood with subsequent platinum / palladium coating and SEM analysis. https://www.leica-microsystems.com//science-lab/human-blood-cells-protocol/ Tue, 24 May 2016 08:03:00 +0000 Dr. W. H. Mueller https://www.leica-microsystems.com/17971 EM Sample Preparation Application Note for Leica EM TIC 3X - Material Research. Purpose: The fibres are embedded in a soft matrix. That makes it difficult to prepare a cross section. Goal: Cross section of the basalt fibres. https://www.leica-microsystems.com//science-lab/cross-sectioning-of-basalt-fibres/ Fri, 20 May 2016 15:31:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/17945 Digital Microscopy Quality Assurance Quality Control in the electronics and microelectronics industry is confronted with many challenges: A variety of different materials in a sample, some of them metallic and reflective, some matt; a variety of material layers and heights, most often very intricate and hard to discern. Users in microelectronics quality control seek to identify deviations from the standard and improve the production processes so that defects are minimized or even eliminated. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-digital-microscopy-used-in-microelectronics-and-electronics-quality-control/ Thu, 19 May 2016 11:23:00 +0000 PhD James DeRose, Claudia Müller https://www.leica-microsystems.com/17977 Confocal Microscopy Live-Cell Imaging A stem cell's decision to self‐renew or differentiate is thought to critically depend on signaling cues provided by its environment. It is unclear whether stem cells have the intrinsic capacity to control their responsiveness to environmental signals that can be fluctuating and noisy. Using a novel single‐cell microRNA activity reporter, we show that miR‐142 is bimodally expressed in embryonic stem cells, creating two states indistinguishable by pluripotency markers. https://www.leica-microsystems.com//science-lab/the-bimodally-expressed-microrna-mir-142-gates-exit-from-pluripotency/ Fri, 13 May 2016 13:47:00 +0000 https://www.leica-microsystems.com/16064 Super-Resolution DNA origami based nanorulers produced by GATTAquant are common standards to test the achievable spatial resolution of super-resolution microscopes. Recently the nanorulers were used to test the performance of the Leica SR GSD 3D microscope. https://www.leica-microsystems.com//science-lab/quantifying-the-resolution-of-a-leica-sr-gsd-3d-localization-microscopy-system-with-2d-and-3d-nanorulers/ Wed, 11 May 2016 07:32:00 +0000 PhD Carsten Forthmann, PhD Jürgen J. Schmied, PhD Tamara Straube https://www.leica-microsystems.com/17939 Laser Microdissection Tumor development results from mutations in our DNA. For their deeper analysis, cancer researchers have to dissect the relevant tissue areas. Here we report the reason why laser microdissection is a perfect tool for this purpose and how this was taught in the course of a workshop held in Brazil. With the Leica LMD system pure tumor material can be selected and dissected for downstream analysis to ensure 100% pure starting material without any risk of cross contamination with healthy cells. https://www.leica-microsystems.com//science-lab/workflows-protocols-laser-microdissection-for-pathology-and-cancer-research/ Wed, 04 May 2016 11:30:00 +0000 Dr. Falk Schlaudraff, Dr. Christoph Greb https://www.leica-microsystems.com/19427 Digital Microscopy From routine quality assurance to advanced 3D analysis of complex surface structures, there is a digital microscope solution to meet almost any request https://www.leica-microsystems.com//science-lab/get-the-most-out-of-digital-microscopy/ Mon, 02 May 2016 10:36:00 +0000 Clinton Smith, Mario J. Gislao https://www.leica-microsystems.com/17957 Super-Resolution Confocal Microscopy Light Sheet Microscopy Neuroscience Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence imaging (SeeDB2). SeeDB2 matches the refractive indices of fixed tissues to that of immersion oil (1.518), thus minimizing both light scattering and spherical aberrations. https://www.leica-microsystems.com//science-lab/super-resolution-mapping-of-neuronal-circuitry-with-an-index-optimized-clearing-agent/ Wed, 27 Apr 2016 10:07:00 +0000 https://www.leica-microsystems.com/17665 Quality Assurance Digital Microscopy Checking specifications is critical: During the manufacture of auto parts, specifications must be met – whether by the auto parts supplier or the automobile manufacturer. It is important that the parts meet specifications as they are critical for maintaining the performance standards and safe operation of automobiles, trucks, and other vehicles during their lifetimes. https://www.leica-microsystems.com//science-lab/automotive-industry-how-suppliers-and-auto-manufacturers-can-verify-parts-specifications-quickly-and-easily/ Mon, 25 Apr 2016 08:21:00 +0000 PhD James DeRose, André Reinhold, Georg Schlaffer https://www.leica-microsystems.com/17967 Surgical Microscopy Ergonomics In dental medicine, the surgical microscope has become increasingly important for high-quality and successful surgeries, particularly in the field of endodontics. A microscope supports the dentist to conduct micro-invasive surgeries which aim to preserve the tooth substance, conserve the tissue, minimize the risks and reduce healing time. To choose the microscope that best fits the dentist’s needs, it is helpful to know some of the decisive features of a modern dental microscope. https://www.leica-microsystems.com//science-lab/six-features-to-consider-when-choosing-a-dental-microscope/ Fri, 22 Apr 2016 16:22:00 +0000 Marie Stockton https://www.leica-microsystems.com/17472 Basics in Microscopy Widefield Microscopy Most microscopes today are operated with a camera. The characteristics of the camera often decide whether the acquired image will reveal what a researcher wants to see. But when diving into camera terminology, the technical terms can be overwhelming. We have compiled the most important terms with a concise explanation to provide orientation. https://www.leica-microsystems.com//science-lab/definitions-of-basic-technical-terms-for-digital-microscope-cameras-and-image-analysis/ Tue, 19 Apr 2016 11:39:00 +0000 Claudia Müller, Dr. Christoph Greb, Dr. Karin Schwab https://www.leica-microsystems.com/17645 EM Sample Preparation Preparation techniques, like several kinds of coating methods play an important role for high resolution scanning electron microscopy (HR-SEM). Nonconductive sample like biological and synthetic samples have to be prepared with a thin conductive layer to prevent charging. https://www.leica-microsystems.com//science-lab/improvement-of-metallic-thin-films-for-hr-sem-by-using-dc-magnetron-sputter-coater/ Wed, 13 Apr 2016 08:30:00 +0000 Dr. Johannes Rattenberger, DI Alexander Melischnig, DI Dr. Ilse Letofsky-Papst, DI Thomas Ganner, Ing. Hartmuth Schröttner https://www.leica-microsystems.com/17931 Live-Cell Imaging sCMOS cameras are more sensitive and are capable of much higher acquisition speed than cameras with other sensor types. Even though CCD cameras are widely used in live cell imaging and time-lapse recordings, researchers are often concerned that their camera does not detect faint signals. In this interview, Dr. Karin Schwab, Product Manager at Leica Microsystems, talks about the characteristics of sCMOS cameras and how researchers benefit from the latest camera sensor technology. https://www.leica-microsystems.com//science-lab/what-makes-scmos-microscope-cameras-so-popular/ Mon, 11 Apr 2016 10:22:00 +0000 Claudia Müller, Dr. Karin Schwab https://www.leica-microsystems.com/17681 Surface Metrology The ecology of plant-eating (herbivorous) mammals is explored through dietary behavior. The Dental Microwear Texture Analysis (DMTA) is a method used to produce a set of dental data to reconstruct dietary habits. Scientists who study the fossil remains of animals (paleontologists) often try to identify the underlying mechanisms driving evolution. Frequently ecological and environmental models are reconstructed with this fossil data. https://www.leica-microsystems.com//science-lab/characterizing-the-evolution-of-plant-eating-herbivorous-mammals-using-dental-tribology/ Thu, 07 Apr 2016 13:02:00 +0000 PhD Gildas Merceron, PhD Cécile Blondel, PhD James DeRose https://www.leica-microsystems.com/17673 Confocal Microscopy Super-Resolution Since the invention of the microscope, there has been continual discussion about the possibility of showing more detailed features of specimens as compared to just magnifying them. In this article we describe the HyVolution concept and how the combination of confocal multiparameter fluorescence imaging at the confocal super-resolution regime with psf-based real deconvolution allows high-speed multicolor imaging with a resolution down to 140 nm. https://www.leica-microsystems.com//science-lab/hyvolution-super-resolution-imaging-with-a-confocal-microscope/ Fri, 01 Apr 2016 07:19:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Constantin Kappel https://www.leica-microsystems.com/17780 Surgical Microscopy Typical ophthalmic procedures require specific levels of light, focus and magnification in each phase of surgery. Changing quickly between these settings is the prerequisite for an uninterrupted and smooth workflow where the surgeon can concentrate completely on his patient. The surgical microscope for ophthalmology Proveo 8 features a unique level of customization that allows to program complete surgical phases and single parameters. Dr. Devesh Varma is one of the first ophthalmologists having tested the Proveo 8 microscope. https://www.leica-microsystems.com//science-lab/how-a-state-of-the-art-microscope-helps-to-optimize-the-workflow-in-cataract-surgery/ Wed, 30 Mar 2016 12:51:00 +0000 Dr. Devesh Varma https://www.leica-microsystems.com/17667 Live-Cell Imaging Widefield Microscopy In order to successfully work with mammalian cell lines, they must be grown under controlled conditions and require their own specific growth medium. In addition, to guarantee consistency their growth must be monitored at regular intervals. This article describes a typical workflow for subculturing an adherent cell line with detailed illustrations of all of the necessary steps. https://www.leica-microsystems.com//science-lab/how-to-do-a-proper-cell-culture-quick-check/ Thu, 24 Mar 2016 09:18:00 +0000 PhD Tamara Straube, Claudia Müller https://www.leica-microsystems.com/17522 Forensics Digital Microscopy Forensic experts work with a broad range of microscopes to examine evidence from firearms and tool marks, documents, forensic or legal medicine, hair and fibers as well as glass and paint. Digital microscopes support many of these investigations, in which the materials can be very different in character. This image gallery shows examples what digital microscopy can reveal in typical forensics’ samples. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-forensics/image-gallery-digital-microscopy-in-forensics-examples-from-ballistics-document-and-trace-evidence-investigations/ Mon, 21 Mar 2016 18:00:00 +0000 Claudia Müller, MSc Michael Doppler https://www.leica-microsystems.com/17669 Confocal Microscopy Super-Resolution Super-resolution refers to any device or method that can resolve better than the classical Abbe limit. Apart from infinite super-resolution techniques such as STED (stimulated emission depletion) and SMLM (single-molecule localization methods) that can theoretically resolve to any detail, there are also methods for limited super-resolution. Here we present HyVolution by Leica, which merges optical super-resolution and computational super-resolution. The optical part is provided by confocal microscopy, and the computational part by deconvolution. Lateral resolution of 140 nm is demonstrated. HyVolution offers multiple fluorescence recording in truly simultaneous mode. https://www.leica-microsystems.com//science-lab/hyvolution-the-smart-path-to-confocal-super-resolution/ Fri, 18 Mar 2016 14:49:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Constantin Kappel https://www.leica-microsystems.com/17636 Widefield Microscopy Basics in Microscopy “Infinity Optics” refers to the concept of a beam path with parallel rays between the objective and the tube lens of a microscope. Flat optical components can be brought into this “Infinity Space” without influencing image formation, which is critical for the utilization of contrast methods such as DIC or fluorescence. Modern microscopy techniques require the addition of multiple optical instruments, such as light sources or laser devices, into the infinite light path. Different approaches to fulfill this need have emerged and are described here. https://www.leica-microsystems.com//science-lab/infinity-optical-systems/ Fri, 11 Mar 2016 16:28:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/17634 Surgical Microscopy 3D visualization of microscopic images not only offers advantages for neurosurgery and ophthalmology, but also provides important benefits for ENT surgery and training. In this interview, Prof. Erwin Offeciers talks about his experiences with 3D visualization in ENT surgery. He is head of the European Institute for Otorhinolaryngology - Head & Neck Surgery of the Sint-Augustinus Hospital in Antwerp, Belgium, and manages Skillslab, the hospital’s own training center for head and neck surgery, neurosurgery and obstetrics & gynaecology. https://www.leica-microsystems.com//science-lab/3d-visualization-in-ent-surgery/ Tue, 08 Mar 2016 11:42:00 +0000 Prof. Erwin Offeciers https://www.leica-microsystems.com/17377 EM Sample Preparation Decoration of grain boundaries in polycrystalline rocks has a long tradition in Structural Geology as in a monomineralic rock the recrystallized grain size is a good indicator for the paleostress conditions. Understanding the mechanical properties of rock salt and its deformation behavior is of major importance for the prediction of long-term stability of nuclear waste repositories, and our understanding of the dynamics of salt-related sedimentary basins which host the majority of oil and gas accumulations on Earth. https://www.leica-microsystems.com//science-lab/triple-beam-ar-ion-milling-with-a-rotary-stage-to-decorate-grain-boundaries-and-substructures-in-rock-salt/ Thu, 03 Mar 2016 11:01:00 +0000 PhD Joyce Schmatz, PhD Oliver Schenk, PhD Jop Klaver, PhD Janos Urai, PhD Wolfgang Grünewald https://www.leica-microsystems.com/17382 CLEM Pathogenic bacteria have developed intriguing strategies to establish and promote infections in their respective hosts. Most bacterial pathogens initiate infectious diseases by adhering to host cells surface. Knowledge of the interplay between the pathogenic organism and the host cells can provide fundamental new insight into the underlying mechanisms of the infectious process and therefore of the disease. Various microscopy techniques have proven to be critical tools to study these events. https://www.leica-microsystems.com//science-lab/imaging-of-host-cell-bacteria-interactions-using-correlative-microscopy-under-cryo-conditions/ Tue, 01 Mar 2016 11:34:00 +0000 Martin Strnad https://www.leica-microsystems.com/17611 Super-Resolution Multiphoton Microscopy Live-Cell Imaging Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level, 3D nanoscale imaging of living cells becomes possible. Our method thus offers robust access to densely packed, axially extended cellular regions that have been notoriously difficult to super-resolve. Our approach also entails a fluorescence read-out scheme that translates molecular sensitivity to local off-switching rates into improved signal-to-noise ratio and resolution. https://www.leica-microsystems.com//science-lab/4pi-resolft-nanoscopy/ Fri, 26 Feb 2016 12:41:00 +0000 https://www.leica-microsystems.com/17559 Education Confocal Microscopy EM Sample Preparation Fluorescence Microscopy MyScope is an online educational site that provides modules on scanning and transmission electron microscopy, scanning probe and atomic force microscopy, confocal microscopy, X-ray diffraction and microanalysis. These advanced research techniques are important in characterization of materials and understanding biological processes. Developed by technique experts across the Australian Microscopy and Microanalysis Research Facility (AMMRF), quality and usability are assured. https://www.leica-microsystems.com//science-lab/good-to-know/myscope-for-global-research-training/ Tue, 23 Feb 2016 19:17:00 +0000 Dr. Bronwen Cribb, Dr. Miles Apperley, Dr. Jenny Whiting https://www.leica-microsystems.com/17444 Digital Microscopy Digital microscopes can be a great help in life science applications such as the documentation in botany, entomology studies and crop science, or the digitization of museum collections. The image quality digital microscopes yield, is stunning – this image gallery gives you an idea. Enjoy! https://www.leica-microsystems.com//science-lab/galleries/image-gallery-digital-microscopy/image-gallery-life-science-imaging-with-the-leica-dvm6-digital-microscope/ Mon, 22 Feb 2016 15:42:00 +0000 PhD James DeRose, PhD Heinrich Bürgers https://www.leica-microsystems.com/17545 Surgical Microscopy In cataract surgery, ophthalmologists rely on the red reflex which provides ideal contrast to visualize the capsule, lens and anterior chamber structure. A new illumination technology in the latest of ophthalmic microscopes now appears to provide great breadth of red reflex enhancement throughout the entire procedure. Dr. Ike Ahmed is one of the first ophthalmologists having tested the Proveo 8 microscope which features the CoAx4 illumination. https://www.leica-microsystems.com//science-lab/the-impact-of-a-stable-red-reflex-and-enhanced-focus-in-cataract-surgery/ Mon, 15 Feb 2016 14:17:00 +0000 Dr. Ike Ahmed https://www.leica-microsystems.com/17543 Surgical Microscopy A stable red reflex is one of the most important features of an ophthalmic surgical microscope for cataract surgery. It’s the red reflex that makes the structure of the lens visible and thus makes for a uncompromised view for a successful and secure surgery. However, conventional red reflex illumination often decreases during the critical phases of the procedure like phacoemulsification. A new illumination technology with four individual beam paths overcomes these drawbacks. https://www.leica-microsystems.com//science-lab/cataract-surgery-with-coax4-illumination/ Tue, 09 Feb 2016 15:31:00 +0000 Manfred Kuster https://www.leica-microsystems.com/16589 EM Sample Preparation HIV-1 undergoes a two-step assembly process. First, an immature noninfectious particle is assembled, which leaves the infected cell. Second, the structural protein, Gag, is cleaved in the virus by the viral protease, and this leads to formation of the infectious virus. We have assembled part of HIV-1 Gag in vitro to form immature virus-like tubular protein arrays, and have solved a subnanometer-resolution structure of these arrays by using cryo-EM and tomography. This structure reveals interactions of the C-terminal capsid domain of Gag that are critical for HIV-1 assembly. https://www.leica-microsystems.com//science-lab/cryo-electron-microscopy-of-tubular-arrays-of-hiv-1-gag-resolves-structures-essential-for-immature-virus-assembly/ Tue, 09 Feb 2016 12:27:00 +0000 https://www.leica-microsystems.com/15896 Stereo Microscopy Fluorescence Microscopy Transparent zebrafish embryos have proved useful model hosts to visualize and functionally study interactions between innate immune cells and intracellular bacterial pathogens, such as Salmonella typhimurium and Mycobacterium marinum. Micro-injection of bacteria and multi-color fluorescence imaging are essential techniques involved in the application of zebrafish embryo infection models. https://www.leica-microsystems.com//science-lab/infection-of-zebrafish-embryos-with-intracellular-bacterial-pathogens/ Fri, 05 Feb 2016 13:35:00 +0000 https://www.leica-microsystems.com/17468 Basics in Microscopy Widefield Microscopy A significant majority of modern optical microscopy techniques require the use of a digital camera. By working with digital devices researchers can observe specimens on a screen in real time or acquire and store images and quantifiable data. Here we introduce the basic principles behind digital camera technologies commonly encountered in scientific imaging. https://www.leica-microsystems.com//science-lab/introduction-to-digital-camera-technology/ Tue, 02 Feb 2016 16:40:00 +0000 Dr. Christoph Greb, Dr. Peter Laskey, Dr. Karin Schwab https://www.leica-microsystems.com/17507 Super-Resolution Previously, the preferred way to study the subtlest elements of the kidney, such as foot processes and the slit diaphragm has been by the use of electron microscopy. Using STED microscopy, we show that the nanoscale localization of slit diaphragm proteins can now be resolved using light microscopy. Even if the nanoscopic resolution has been available for a decade, light microscopy studies of the slit diaphragm are not found in the literature. This is likely due to the difficulties of achieving the high quality of fluorescent labelling needed for super-resolution microscopy. By applying an optical clearing protocol based on the CLARITY technique, we found that the immunostaining quality in kidney tissue can be improved. The improvement is likely due to the removal of lipids, resulting in a higher availability of binding epitopes in cleared tissue, as compared to PFA fixed non-cleared tissue. https://www.leica-microsystems.com//science-lab/how-to-combine-sted-and-clarity/ Fri, 29 Jan 2016 14:46:00 +0000 https://www.leica-microsystems.com/13262 Quantitative Imaging Fast FRAP experiments need a sufficient number of measurement points for meaningful interpretation and fitting analysis. To study very fast translocational processes, the use of a resonant scanner (RS) is preferred. The advantage in using FRAP with the RS is that statistics are much better in experiments that require fast acquisition: If the half time of recovery is about 0.5 sec you may have only about 3 to 4 data points using the conventional scanner, whereas with the resonant scanner you can get about 20 data points. https://www.leica-microsystems.com//science-lab/frap-with-tcs-sp8-resonant-scanner/ Tue, 26 Jan 2016 15:39:00 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/17437 Fluorescence Microscopy Stereo Microscopy Over their entire lifetime, 1 out of 8 women can suffer from breast cancer. The risk of developing breast cancer (mammary carcinogenesis) increases with a woman’s age and is related to her reproductive history. The chance of mammary carcinogenesis is less for women who give birth to a child before the age of 30. However, it is now known that the risk of breast cancer also can depend on the number of times a woman experiences elevated blood serum progesterone levels, in relation to the menstrual cycle, before her first pregnancy. https://www.leica-microsystems.com//science-lab/investigating-the-influence-of-hormone-signaling-pathways-on-mammary-gland-development-and-the-onset-of-breast-cancer/ Thu, 21 Jan 2016 11:12:00 +0000 MD Cathrin Brisken, PhD Renuga Devi Rajaram, PhD Duje Buric, MSc Ayyakkannu Ayyanan, PhD James DeRose https://www.leica-microsystems.com/17418 Surface Metrology Optical Coherence Tomography Although interferometry may once have been dismissed by the medical community because of its inability to see inside the human body, its application within the wider biomedical arena is continuing to grow and evolve; so much so that the technique is now used in the production of prosthetic limbs, research in dentistry and even for cultivating stem cells. https://www.leica-microsystems.com//science-lab/a-healthy-dose-of-interferometry/ Thu, 14 Jan 2016 15:06:00 +0000 https://www.leica-microsystems.com/17398 Quality Assurance Digital Microscopy For the past several years, digital microscopy has been shown to be useful for inspection, quality control and assurance (QC/QA), and failure analysis (FA) in the microelectronics industry, especially for printed circuit boards (PCBs). Recently, state-of-the-art improvements have made digital microscopy even more powerful and practical for inspection, leading to a more efficient workflow. Here, the advantages of certain digital microscope features, i.e., intuitive software for operation and analysis, fast and easy ways to change magnification, and encoding for reliable recall of parameters, are explained. https://www.leica-microsystems.com//science-lab/inspecting-and-analyzing-printed-circuit-boards-quickly-and-reliably-with-a-digital-microscope/ Mon, 11 Jan 2016 09:09:00 +0000 PhD James DeRose, Georg Schlaffer https://www.leica-microsystems.com/16796 Super-Resolution How to break a fundamental law of physics and win a Nobel Prize to boot. Stefan Hell explains super-resolved fluorescence microscopy for which he shared the 2014 Nobel Prize in chemistry. https://www.leica-microsystems.com//science-lab/video-fluorescence-is-a-state-of-mind/ Thu, 07 Jan 2016 20:45:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/17379 EM Sample Preparation Sun screen lotion was carefully filled in the 100 μm incision of a 3 mm copper/gold plated flat carrier and covered with 3 mm sapphire disk. The sun screen lotion sample was then high pressure frozen with a Leica EM ICE with and subsequently without light stimulation. The light stimulated samples were exposed to a UV light for 500 milliseconds. https://www.leica-microsystems.com//science-lab/high-pressure-freezing-with-light-stimulation/ Fri, 18 Dec 2015 12:01:00 +0000 PhD Dietmar Pum, Dr. Cveta Tomova, PhD Saskia Mimietz-Oeckler https://www.leica-microsystems.com/17358 EM Sample Preparation Quantitative and analytical analysis at high spatial resolution places stringent demands on the quality of the produced TEM specimens. Pristine and high-quality samples are indispensible for atomic resolution TEM analysis. In this application note a general procedure for obtaining cross-sectional and plan-view TEM specimens using the Leica EM RES102 ion milling system is outlined. The procedure described below can be easily adapted for a large range of materials e.g. thin film materials, semiconductors, multilayered materials, ceramics, superconductors, … https://www.leica-microsystems.com//science-lab/tem-sample-preparation-made-easy-prepare-tem-specimen-by-broad-beam-argon-ion-milling/ Tue, 15 Dec 2015 12:03:00 +0000 PhD Frédéric Leroux https://www.leica-microsystems.com/16593 Super-Resolution Neuroscience At the presynaptic active zone (AZ) a variety of specialized proteins are assembled to complex architectures, which set the basis for speed, precision and plasticity of synaptic transmission. Recently, super-resolution microscopy (SRM) techniques have begun to enter the neurosciences. These approaches combine high spatial resolution with the molecular specificity of fluorescence microscopy. Here, we discuss how SRM techniques can be used to obtain information on the organization of AZ proteins. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-of-the-synaptic-active-zone/ Tue, 15 Dec 2015 10:15:00 +0000 Nadine Ehmann https://www.leica-microsystems.com/16591 Super-Resolution Natural killer (NK) cells are cytolytic effector cells of the innate immune system. Here, we show that deconstruction of synaptic cortical filamentous (F)-actin by Coronin 1A (Coro1A) is required for NK cell cytotoxicity through the remodeling of F-actin to enable lytic granule secretion. We define this requirement for remodeling using superresolution nanoscopy and Coro1A-deficient NK cells. In addition, we use NK cells from a patient with a rare Coro1A mutation, thus illustrating a critical link between Coro1A function and human health. https://www.leica-microsystems.com//science-lab/lytic-immune-synapse-function-requires-filamentous-actin-deconstruction-by-coronin-1a/ Thu, 10 Dec 2015 10:21:00 +0000 Ph.D. Emily M. Mace, Ph.D., MD Jordan S. Orange https://www.leica-microsystems.com/16814 Laser Microdissection Neuroscience The main topic during this course was how to apply laser microdissection in neuroscience. Leica specialists demonstrated why laser microdissection is a suitable techniques for brain investigation as it allows to separate distinct brain layers or even to isolate individual neurons. https://www.leica-microsystems.com//science-lab/workflows-protocols-connecting-microscopy-and-molecular-biology-in-neuroscience/ Fri, 04 Dec 2015 16:01:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/16808 Laser Microdissection During the first Leica Workshop in Brazil, at the Centro de Energia Nuclear na Agricultura/USP (CENA), the participants learned how to prepare samples for laser microdissection (LMD) using a cryotome. Another topic was the dissection of individual chromosomes from chromosome spreads. Leica specialists held a short training session with the LMD. After this, new LMD users were able to run the system and practiced how to dissect chromosomes and collect single chromosomes for downstream analysis. https://www.leica-microsystems.com//science-lab/workflows-protocols-how-to-isolate-individual-chromosomes-with-laser-microdissection/ Thu, 03 Dec 2015 20:06:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/16798 Basics in Microscopy Stereo Microscopy Widefield Microscopy There are many influences on color in the imaging process including lighting, optics, sensor, and monitor, and ultimately print. The first, and generally most important, is lighting. There are plenty of options for light sources, Halogen, LED, and arc lamps are among the most popular for microscopes. Each light source has its own advantages and disadvantages and it is up to the user to learn which is best for the sample and application. https://www.leica-microsystems.com//science-lab/color-infidelity-why-using-a-light-source-incorrectly-is-cheating-on-your-data/ Thu, 19 Nov 2015 16:28:00 +0000 Mike Tjepkema https://www.leica-microsystems.com/16565 Super-Resolution Scientists from three Max Planck Institutes have gained fundamental insights into the organization of mitochondrial DNA (mtDNA). The researchers observed in high-resolution images gained with nobel prize-winning microscopy techniques that single copies of mtDNA are packaged by a specialized protein into slightly elongated structures of circa 100 nm in length. https://www.leica-microsystems.com//science-lab/cross-strand-binding-of-tfam-to-a-single-mtdna-molecule-forms-the-mitochondrial-nucleoid/ Tue, 17 Nov 2015 17:20:00 +0000 Dr. Christian Kukat https://www.leica-microsystems.com/16546 Optical Coherence Tomography The role of iOCT in vitreoretinal surgery continues to be defined by active research and enhancements to integrative technologies. Further research is needed to better define the specific applications of iOCT that impact patient outcomes and surgical decision-making. Future advancements in integrative systems, OCT-friendly instrumentation, and software algorithms will further expand the horizon of iOCT in the vitreoretinal surgical theater. As OCT transformed the clinical management of the vitreoretinal conditions, iOCT has the potential to be a paradigm-shifting technology in the operating room. https://www.leica-microsystems.com//science-lab/the-value-of-intraoperative-oct-imaging-in-vitreoretinal-surgery/ Mon, 09 Nov 2015 13:15:00 +0000 https://www.leica-microsystems.com/16544 Optical Coherence Tomography Monitoring young children with optic pathway gliomas (OPGs) for visual deterioration can be difficult owing to age-related noncompliance. Optical coherence tomography (OCT) measures of retinal nerve fiber layer (RNFL) thickness have been proposed as a surrogate marker of vision but this technique is also limited by patient cooperation. To determine whether measures of circumpapillary RNFL thickness, acquired with handheld OCT (HH-OCT) during sedation, can differentiate between young children with and without vision loss from OPGs. https://www.leica-microsystems.com//science-lab/handheld-optical-coherence-tomography-during-sedation-in-young-children-with-optic-pathway-gliomas/ Mon, 09 Nov 2015 13:13:00 +0000 https://www.leica-microsystems.com/16542 Optical Coherence Tomography This study assessed the feasibility of High Resolution Spectral Domain Optical Coherence Tomography (HR-SDOCT) to guide donor tissue preparation in Descemet's Membrane Endothelial Keratoplasty (DMEK) using the reverse big bubble technique. https://www.leica-microsystems.com//science-lab/high-resolution-optical-coherence-tomography-guided-donor-tissue-preparation-for-descemets-membrane-endothelial-keratoplasty-using-reverse-big-bubble-technique/ Mon, 09 Nov 2015 13:12:00 +0000 https://www.leica-microsystems.com/16540 Optical Coherence Tomography OCT is a non-invasive, contact-free technique that helps to detect and monitor morphological changes of ocular tissue, in particular retinal layer thickness, which can give insight into pathological conditions such as glaucoma, age-related macular degeneration (AMD) or diabetic retinopathy. This references list presents selected publications performed with an Envisu OCT imaging system. https://www.leica-microsystems.com//science-lab/oct-publication-list/ Mon, 09 Nov 2015 13:10:00 +0000 Eric Buckland https://www.leica-microsystems.com/16532 Optical Coherence Tomography Surgical Microscopy The high-resolution, cross sectional information provided by iOCT creates a unique feedback system for vitreoretinal surgery. The PIONEER Study conducted by the Cole Eye Institute is a three-year exploratory study to investigate clinical feasibility, the impact on surgical decision-making, and the potential safety of a microscope-mounted portable iOCT. PIONEER represents the largest study to date examining the feasibility, utility, and safety of iOCT. https://www.leica-microsystems.com//science-lab/intraoperative-oct-today-and-tomorrow-the-pioneer-study-two-year-results-and-beyond/ Mon, 09 Nov 2015 13:09:00 +0000 https://www.leica-microsystems.com/16536 Optical Coherence Tomography Surgical Microscopy The work prepared and presented within this video demonstrates preliminary research, and the promising future of perioperative OCT to aid the diagnosis of physiological and pathological conditions during ophthalmic surgery in order to help guide surgical decision making in anterior segment cases. Showing SDOCT Images acquired with an Envisu SD-OCT system, Prof. Yoo demonstrates the use of interoperative OCT imaging several important surgical procedures such as DALK, DSAEK and DMEK. https://www.leica-microsystems.com//science-lab/video-talk-by-sonia-h-yoo-interoperative-oct-imaging-of-selective-lamellar-keratoplasty/ Mon, 09 Nov 2015 13:07:00 +0000 Prof. Sonia H. Yoo https://www.leica-microsystems.com/16790 Surgical Microscopy The concept of minimal intervention dentistry became a primary approach to treat carious lesions. These days, this concept could be adapted to manage soft tissue around the teeth and implants as a minimally invasive technique. This video, we show a case of traumatic soft and hard tissue wound on the maxillary anterior and its rehabilitation. The patient was a 22 year-old women, and her chief complaint was severe aesthetic disorder because of her trauma. She lost two incisor teeth as well as tissue and bone around the teeth. https://www.leica-microsystems.com//science-lab/video-traumatic-soft-and-hard-tissue-wound-on-the-maxillary-anterior-with-microscope-assisted-rehabilitation/ Mon, 02 Nov 2015 08:02:00 +0000 DDS, PhD Takuya Satoh https://www.leica-microsystems.com/16804 EM Sample Preparation In 2014, the renowned Electron Microscopy for Materials Science (EMAT) research lab at the University Antwerp, Belgium, and Leica Microsystems started a fruitful collaboration to establish a Leica Reference Site in Antwerp. This site, officially opened in July 2014, is dedicated to specimen preparation for electron microscopy in materials science with a special focus on ion beam milling and recently also on carbon coating. In this interview Prof Gustaf van Tendeloo, Director of EMAT, and Frédéric Leroux, TEM specimen preparation specialist, talk about research topics at EMAT, how the Leica reference site has evolved, and future trends for EM sample preparation. https://www.leica-microsystems.com//science-lab/electron-microscopy-sample-preparation-the-future-is-cold-dynamic-and-hybrid/ Thu, 22 Oct 2015 19:22:00 +0000 Prof. Gustaaf (Staf) Van Tendeloo, PhD Frédéric Leroux, Florence Hauger https://www.leica-microsystems.com/16800 Ergonomics Stereo Microscopy The thought of a Drosophoila headed to the local bar for a drink creates a funny joke, but after long hours of sorting flies under a microscope that causes eyestrain or neck pain, you may be the one that wants to head to the local bar for a drink! Unfortunately most users do not know why they experience discomfort when using a stereo microscope. https://www.leica-microsystems.com//science-lab/ergonomics/what-do-you-call-a-drosophila-who-likes-to-drink-a-bar-fly/ Wed, 21 Oct 2015 16:22:00 +0000 Mike Tjepkema https://www.leica-microsystems.com/16785 Stereo Microscopy Quality Assurance This report gives users of stereo microscopes helpful advice when attempting to select optimal illumination or lighting systems for sample observation. The illumination used for microscopic observation has a very important effect on the final image quality. Choosing the illumination to achieve the best results depends upon the type of sample and its features of interest, as well as the application and purpose for microscopic observation. The following information should help microscope users to choose illumination systems that produce the best imaging results. https://www.leica-microsystems.com//science-lab/illumination-lighting-systems-for-stereo-microscopes/ Tue, 20 Oct 2015 08:51:00 +0000 PhD James DeRose, Matthias Schacht https://www.leica-microsystems.com/15702 Stereo Microscopy We describe a new extinct lineage Manipulatoridae (new family) of cockroaches from the Upper Cretaceous (Cenomanian) amber of Myanmar. Manipulator modificaputis gen. et sp. n. is a morphologically unique extinct cockroach that represents the first (of a total of 29 known worldwide) cockroach family reported exclusively from the Myanmar amber. https://www.leica-microsystems.com//science-lab/new-predatory-cockroaches-insecta-blattaria-manipulatoridae-famn-from-the-upper-cretaceous-myanmar-amber/ Wed, 14 Oct 2015 16:22:00 +0000 https://www.leica-microsystems.com/13935 Fluorescence Microscopy Live cell imaging has been revolutionized by the discovery of the green fluorescent protein (GFP). This lecture covers the history of GFP, how it folds and becomes fluorescent, how it has been mutated to produce additional colors (blue, cyan, yellow), and the discovery of red fluorescent proteins from corals. It also covers novel photoswitchable and photoactivatible fluorescent proteins, whose color can be changed by light, and new infrared fluorescent proteins. https://www.leica-microsystems.com//science-lab/video-talk-by-roger-tsien-fluorescent-proteins/ Wed, 07 Oct 2015 16:55:00 +0000 PhD Roger Y. Tsien https://www.leica-microsystems.com/16587 Light Sheet Microscopy Live-Cell Imaging Confocal Microscopy Living cells and organisms often suffer from the high light intensities used for fluorescent imaging. Light sheet microscopy reduces phototoxic effects and bleaching by illuminating a specimen in only a single plane at a time. A new light sheet microscope combines light sheet and confocal microscopy in one system without compromising either functionality and allows the combination of the two methods, e.g. confocal photomanipulation with subsequent light sheet acquisition, for new applications. https://www.leica-microsystems.com//science-lab/light-sheet-microscopy-turned-vertically/ Fri, 02 Oct 2015 13:34:00 +0000 PhD Isabelle Köster, Dr. Petra Haas https://www.leica-microsystems.com/16458 Light Sheet Microscopy Super-Resolution Confocal Microscopy Bram van den Broek is a postdoctoral fellow at the Netherlands cancer institute in Amsterdam where he supports the advanced microscopy techniques in the laboratory of Kees Jalink. Working with Leica Microsystems as a collaboration partner for beta-testing of microscopes he enjoys very much. https://www.leica-microsystems.com//science-lab/the-leica-digital-light-sheet-module-a-clever-example-of-thinking-out-of-the-box/ Tue, 29 Sep 2015 14:39:00 +0000 Bram van den Broek, Dipl. oec.-troph. Anja Schué, PhD Isabelle Köster https://www.leica-microsystems.com/16512 Confocal Microscopy Contact-dependent intercellular transfer (codeIT) of cellular constituents can have functional consequences for recipient cells, such as enhanced survival and drug resistance. Here, we present a novel microscopy-based screening method to identify regulators and cargo of codeIT. Single donor cells, carrying fluorescently labelled endocytic organelles or proteins, are co-cultured with excess acceptor cells. CodeIT is quantified by confocal microscopy and image analysis in 3D, preserving spatial information. An siRNA-based screening using this method revealed the involvement of several myosins and small GTPases as codeIT regulators. https://www.leica-microsystems.com//science-lab/novel-microscopy-based-screening-method-reveals-regulators-of-contact-dependent-intercellular-transfer/ Wed, 23 Sep 2015 09:50:00 +0000 https://www.leica-microsystems.com/16521 Super-Resolution Live-Cell Imaging Fluorescence Microscopy We have been developing new tools based on fluorogenic forced intercalation (FIT) probes for RNA detection quantification and interference in biological samples. Upon duplex formation with target nucleic acids, the base surrogates TO dye increases its quantum yield and brightness substantially (>10 fold). https://www.leica-microsystems.com//science-lab/probes-that-fit-rna/ Tue, 22 Sep 2015 17:25:00 +0000 PhD Imre Gaspar https://www.leica-microsystems.com/16035 Fluorescence Microscopy Live-Cell Imaging Basics in Microscopy Green fluorescent protein (GFP) isolated from the jellyfish Aequorea victoria and GFP-like fluorescent proteins from other animals have had an important role in the technical innovations that have driven these advances. This poster provides a comprehensive user's guide to fluorescent proteins and sensors , their key properties and the cell biological questions to which they can be applied. https://www.leica-microsystems.com//science-lab/fluorescent-proteins-illuminate-cell-biology/ Fri, 18 Sep 2015 09:03:00 +0000 https://www.leica-microsystems.com/18917 Super-Resolution TIRF Microscopy Co-localization analysis is a widely used tool to seek evidence for functional interactions between molecules in different color channels in microscopic images. Here we extend the basic co-localization analysis by including the orientations of the structures on which the molecules reside. We refer to the combination of co-localization of molecules and orientational alignment of the structures on which they reside as co-orientation. Because the orientation varies with the length scale at which it is evaluated, we consider this scale as a separate informative dimension in the analysis. Additionally we introduce a data driven method for testing the statistical significance of the co-orientation and provide a method for visualizing the local co-orientation strength in images. We demonstrate our methods on simulated localization microscopy data of filamentous structures, as well as experimental images of similar structures acquired with localization microscopy in different color channels. https://www.leica-microsystems.com//science-lab/co-orientation-quantifying-simultaneous-co-localization-and-orientational-alignment-of-filaments-in-light-microscopy/ Wed, 16 Sep 2015 15:20:00 +0000 Robert Nieuwenhuizen, MSc Leila Nahidiazar https://www.leica-microsystems.com/16124 Basics in Microscopy The history of microscopy begins in the Middle Ages. As far back as the 11th century, plano-convex lenses made of polished beryl were used in the Arab world as reading stones to magnify manuscripts. However, the further development of these lenses into the first microscopes cannot be attributed to any one person. It took the ideas and designs of many scientists and scholars to produce instruments capable of strong magnification. https://www.leica-microsystems.com//science-lab/a-brief-history-of-light-microscopy-from-the-medieval-reading-stone-to-super-resolution/ Tue, 08 Sep 2015 17:41:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/15426 Confocal Microscopy Collective beta cell activity in islets of Langerhans is critical for the supply of insulin within an organism. In order to get a detailed insight into the functional organization of the syncytium, we applied advanced analytical tools from the realm of complex network theory to uncover the functional connectivity pattern among cells composing the intact islet. https://www.leica-microsystems.com//science-lab/progressive-glucose-stimulation-of-islet-beta-cells-reveals-a-transition-from-segregated-to-integrated-modular-functional-connectivity-patterns/ Thu, 03 Sep 2015 17:10:00 +0000 https://www.leica-microsystems.com/16097 Super-Resolution Here we show staining of HDAC1 in cancer tissue and epidermoid carcinoma cells. These results clearly show that the use of appropriate validated antibodies and STED microscopy are important tools to study subcellular structures beyond the diffraction limit correcting ill-defined images. This is critical in co-localization studies of proteins inside cells. https://www.leica-microsystems.com//science-lab/localization-of-hdac1-using-super-resolution-sted-microscopy/ Wed, 02 Sep 2015 15:57:00 +0000 PhD Karin Abarca Heidemann, PhD Sarah Crowe, MS, PhD Tobias Jacob, PhD Carl A. Ascoli https://www.leica-microsystems.com/16460 Confocal Microscopy Multiphoton Microscopy This article outlines the most important sensors used in confocal microscopy. By confocal microscopy, we mean "True Confocal Scanning", i.e. the technique that illuminates and measures one single point only. The aim is not to impart in-depth specialist knowledge, but to give the user a small but clear overview of the differences between the various technologies and to advise on which sensor may be most suitable for which applications. https://www.leica-microsystems.com//science-lab/from-light-to-mind-sensors-and-measuring-techniques-in-confocal-microscopy/ Fri, 28 Aug 2015 16:25:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/15200 Confocal Microscopy The Leica White Light Laser produces a continuous spectral output between the wavelengths of 470 and 670 nm. It allows you to select 8 excitation lines from 3 trillion unique combinations for simultaneous imaging of multiple fluorophores. The white light laser source of the Leica TCS SP8 X perfectly matches the wavelength of any fluorophore. https://www.leica-microsystems.com//science-lab/video-the-white-light-laser-how-to-effectively-excite-multiple-fluorophores-with-a-single-light-source/ Thu, 27 Aug 2015 10:05:00 +0000 PhD Christopher Vega https://www.leica-microsystems.com/15928 Super-Resolution The maximization of the useful (within the time gate) photon flux is then an important aspect to obtain super-resolved STED images. Here we show that by using a fast-gated single-photon avalanche diode (SPAD), i.e. a detector able to rapidly (hundreds picoseconds) switch-on and -off can improve significantly the signal-to-noise ratio (SNR) of the gated STED image. In addition to an enhancement of the image SNR, the use of the fast-gated SPAD reduces also the system complexity. We demonstrate these abilities both on calibration and biological sample. https://www.leica-microsystems.com//science-lab/gated-sted-microscopy-with-time-gated-single-photon-avalanche-diode/ Tue, 25 Aug 2015 17:24:00 +0000 https://www.leica-microsystems.com/16095 Stereo Microscopy For samples/specimens immersed in a liquid or embedded in a polymer, high quality microscopic observation can be hindered as a result of spherical aberration. An objective which can correct for refractive index mismatch allows images with greatly reduced spherical aberration and sharper focus to be obtained. https://www.leica-microsystems.com//science-lab/how-to-correct-aberration-in-stereo-microscopy-by-using-the-right-objective-lenses/ Fri, 21 Aug 2015 17:27:00 +0000 PhD James DeRose, PhD Heinrich Bürgers https://www.leica-microsystems.com/16099 Confocal Microscopy Jutta Maria Bulkescher is the technical coordinator in the Novo Nordisk Foundation Center for Protein Research and Danish Stem Cell Center in Copenhagen, Denmark. The Leica HCS-A matrix screener is an invaluable tool for her facility. "It just gives us the biggest and easiest flexibility we can have to set up different imaging paramters and to check different conditions on one multi-well plate", explains Bulkescher. https://www.leica-microsystems.com//science-lab/we-can-go-home-and-the-imaging-is-done-automatically-by-the-leica-hcs-a-matrix-screener/ Wed, 19 Aug 2015 14:30:00 +0000 Jutta Bulkescher, PhD Isabelle Köster, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/16033 Super-Resolution The widely popular class of quantum-dot molecular labels could so far not be utilized as standard fluorescent probes in STED (stimulated emission depletion) nanoscopy. This is because broad quantum-dot excitation spectra extend deeply into the spectral bands used for STED, thus compromising the transient fluorescence silencing required for attaining super-resolution. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-with-fluorescent-quantum-dots/ Thu, 13 Aug 2015 09:24:00 +0000 https://www.leica-microsystems.com/14400 Basics in Microscopy Joseph Gall takes us through the history of early microscopes and the discovery of the cell. Compound microscopes were invented alongside the telescope in the 17th century; however these microscopes were not widely used until the late 19th century due to optical aberrations. In the meantime, simple microscopes were used throughout the 1700s and 1800s to make major discoveries in biology, including the first descriptions of the nucleus, cilia, cells, bacteria, and protozoans. Once optics improved in the mid to late 1800s, compound microscopes were used to discover chromosomes, mitosis, and other cellular structures. https://www.leica-microsystems.com//science-lab/video-talk-by-joseph-gall-early-history-of-microscopy/ Tue, 11 Aug 2015 07:48:00 +0000 BS, PhD Joseph Gall https://www.leica-microsystems.com/16165 EM Sample Preparation In June 2014, the Institut de Minéralogie, de Physique des Matériaux et de Cosmochimie (IMPMC) of the Université Pierre et Marie Curie in Paris became a Leica reference lab for EM Sample Preparation with equipment like the Leica EM TXP target surfacing systems and the Leica EM TIC 3X ion beam milling system. In the interview, Imène Estève, engineer at the Centre National de Recherches Scientifiques (CNRS) and head of the SEM-FIB national facility, in charge of running the lab, tells us about how this cooperation has developed. https://www.leica-microsystems.com//science-lab/a-good-place-for-materials-scientists-and-mineralogists-to-prepare-their-em-samples/ Fri, 07 Aug 2015 09:31:00 +0000 MPhys, MChem Imène Esteve, Florence Hauger, Claudia Müller https://www.leica-microsystems.com/16093 Education Basics in Microscopy Microscopes and other optical instruments can be affected during use by environmental factors. The environment depends on the geographic location and conditions of the place where the instrument is put to use. Usually, microscopes are manufactured in a way to ensure instrument robustness to a variety of conditions. https://www.leica-microsystems.com//science-lab/microscopes-put-to-the-test-with-severe-conditions-iso-standard-for-resistance-of-optical-instruments-to-fungus-and-mold-growth/ Wed, 05 Aug 2015 07:15:00 +0000 PhD James DeRose, Vince Vaccarelli https://www.leica-microsystems.com/16176 Super-Resolution Quantitative Imaging Recent years have seen the development of multiple technologies to investigate, with great spatial and temporal resolution, the dynamics of lipids in cellular and model membranes. One of these approaches is the combination of far-field super-resolution stimulated-emission-depletion (STED) microscopy with fluorescence correlation spectroscopy (FCS). STED-FCS combines the diffraction-unlimited spatial resolution of STED microscopy with the statistical accuracy of FCS to determine sub-millisecond-fast molecular dynamics with single-molecule sensitivity. https://www.leica-microsystems.com//science-lab/a-straightforward-approach-for-gated-sted-fcs-to-investigate-lipid-membrane-dynamics/ Mon, 03 Aug 2015 10:29:00 +0000 https://www.leica-microsystems.com/16386 Surgical Microscopy Surgeons at The Children’s Hospital of Philadelphia (CHOP) joined with colleagues from Penn Medicine recently to complete the world’s first bilateral hand transplant on a child. Earlier this month, the surgical team successfully transplanted donor hands and forearms onto 8-year-old Zion Harvey who, several years earlier, had undergone amputation of his hands and feet and a kidney transplant following a serious infection. https://www.leica-microsystems.com//science-lab/first-bilateral-hand-transplant-in-a-child/ Fri, 31 Jul 2015 11:26:00 +0000 https://www.leica-microsystems.com/16174 Super-Resolution Quantitative Imaging Live-Cell Imaging Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but this concept remains controversial because this phenomenon has thus far only been observed with artefact-prone probes in combination with a single technique: single particle tracking. https://www.leica-microsystems.com//science-lab/cortical-actin-networks-induce-spatio-temporal-confinement-of-phospholipids-in-the-plasma-membrane-a-minimally-invasive-investigation-by-sted-fcs/ Fri, 24 Jul 2015 13:02:00 +0000 Débora Machado Andrade https://www.leica-microsystems.com/15990 Forensics When the first compound microscopes were invented in 1590, scientists marveled at their new ability to see tiny objects and features that were previously invisible to the eye and therefore seemingly nonexistent. Ever since then, the study of these miniscule details has brought science into a forensic world once ruled by intuition and deduction. Choosing a microscope with the right optics can reduce these hidden errors considerably to provide results that are both more accurate and more reproducible – two attributes that are both essential in modern forensics. https://www.leica-microsystems.com//science-lab/using-telecentric-optical-systems-to-optimize-forensic-image-accuracy-and-reproducibility/ Fri, 17 Jul 2015 15:17:00 +0000 Claus Klein, Wayne Buttermore, MSc Michael Doppler https://www.leica-microsystems.com/15239 Super-Resolution Quantitative Imaging The eukaryotic cell membrane is connected to a dense actin rich cortex. We present FCS and STED experiments showing that dense membrane bound actin networks have severe influence on lipid phase separation. Our results reveal a mechanism how cells may prevent macroscopic demixing of their membrane components, while at the same time regulate the local membrane composition. https://www.leica-microsystems.com//science-lab/a-lipid-bound-actin-meshwork-organizes-liquid-phase-separation-in-model-membranes/ Wed, 15 Jul 2015 13:49:00 +0000 https://www.leica-microsystems.com/16110 Laser Microdissection Laser Microdissection (LMD) allows isolating individual cells or chromosomes and is a well established technique for sample preparation prior downstream analysis of the nucleic acid content via PCR or sequencing techniques. Here we describe the successful combination of the Leica Microsystems LMD system and Qiagen kits for purification of nucleic acids even from little amounts. https://www.leica-microsystems.com//science-lab/workflows-protocols-how-to-use-a-leica-laser-microdissection-system-and-qiagen-kits-for-successful-rna-analysis/ Fri, 10 Jul 2015 14:26:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/15241 Super-Resolution Live-Cell Imaging Most proteins have uneven distributions in the plasma membrane. Broadly speaking, this may be caused by mechanisms specific to each protein, or may be a consequence of a general pattern that affects the distribution of all membrane proteins. https://www.leica-microsystems.com//science-lab/multi-protein-assemblies-underlie-the-mesoscale-organization-of-the-plasma-membrane/ Thu, 09 Jul 2015 13:44:00 +0000 https://www.leica-microsystems.com/16010 EM Sample Preparation X-ray micro-computed tomography (micro-CT) is a routinely applied non-invasive technique for the investigation of the internal anatomy and morphology of organisms. As a result of a micro-CT scan a stack of grey-scale images is generated from a series of projections taken at defined angles during sample rotation. Since several years the number of lab-based micro-CT imaging systems is constantly growing making this technique available to a broad spectrum of researchers and applications. https://www.leica-microsystems.com//science-lab/critical-point-drying-for-the-preparation-of-biological-samples-for-microct-analysis/ Mon, 06 Jul 2015 13:23:00 +0000 PhD Peter Michalik, PhD Elisabeth Lipke https://www.leica-microsystems.com/15596 EM Sample Preparation Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challenge because no detailed protocols for ultrastructural analysis of cysts are currently available. https://www.leica-microsystems.com//science-lab/transmission-electron-microscopy-sample-preparation-protocols-for-the-ultrastructural-study-of-cysts-of-free-living-protozoa/ Thu, 02 Jul 2015 17:25:00 +0000 https://www.leica-microsystems.com/15138 Digital Microscopy In recent years, a number of technological advancements started to modify the long standing appearance and functionalities of traditional optical microscopes used in Pathology and other medical fields. In fact, at present many new tools for microscopical visualization exist that are based on digital imaging, robotization, and remote communication. Such tools are typically adopted in activities ranging from education to telediagnosis to remote consultation. https://www.leica-microsystems.com//science-lab/transnational-telepathology-consultations-using-a-basic-digital-microscope-experience-in-the-italy-slovenjia-interreg-project-patient-without-borders/ Mon, 29 Jun 2015 10:22:00 +0000 https://www.leica-microsystems.com/11767 Digital Microscopy When the first multi-lens microscopes were invented in 1590, scientists marveled at their new ability to SEE small objects and features in the natural world that were previously invisible to the eye and therefore seemingly nonexistent. With the constant miniaturization of parts and products in automated manufacturing over the past 5 decades, the use of microscopes has spread increasingly from science to industry. Today microscopes are found in a multitude of assembly and inspection applications wherever visualization and measurement of miniscule features are required. https://www.leica-microsystems.com//science-lab/using-telecentric-optical-systems-to-optimize-industrial-image-accuracy-and-reproducibility/ Thu, 25 Jun 2015 18:57:00 +0000 Clinton Smith https://www.leica-microsystems.com/16019 Super-Resolution The 5th Super-Resolution User Club Meeting was held in collaboration with Professor Kees Jalink and The Netherlands Cancer Institute (NKI) in Amsterdam. Having the meeting at a location where super-resolution microscopy is used on a daily basis makes a big difference, offering participants the chance to use live cells for workshops and see systems working in their true environments. Thanks also to the scientists that supported the meeting by coming and giving talks. As super-resolution continues to grow in importance in research, we recognize the need to come together to network, share information and experiences. Here we present the abstracts of the talks. https://www.leica-microsystems.com//science-lab/abstracts-of-the-5th-european-super-resolution-user-club-meeting/ Tue, 23 Jun 2015 13:23:00 +0000 PhD Kees Jalink, PhD Jürgen J. Schmied, PhD Jonas Ries, PhD David Williamson, MSc Daniela Leyton Puig, Robert Nieuwenhuizen, MSc Leila Nahidiazar, PhD Luc Reymond, PhD Imre Gaspar, Luca Lanzanò, Ivan Michel Antolovic https://www.leica-microsystems.com/15894 Confocal Microscopy Microscopy in the context of biomedical research is demanding new tools to automatically detect and capture objects of interest. The few extant packages addressing this need, however, have enjoyed limited uptake due to complexity of use and installation. To overcome these drawbacks, we developed iMSRC, which combines ease of use and installation with high flexibility and enables applications such as rare event detection and high-resolution tissue sample screening, saving time and resources. https://www.leica-microsystems.com//science-lab/imsrc-converting-a-standard-automated-microscope-into-an-intelligent-screening-platform/ Mon, 22 Jun 2015 11:56:00 +0000 https://www.leica-microsystems.com/15096 Stereo Microscopy We describe a technique of microinjecting the aminoglycoside, gentamicin, into 2 days post-fetilization (dpf) zebrafish larvae to induce acute kidney injury (AKI). We also describe a method for whole mount immunohistochemistry, plastic embedding and sectioning of zebrafish larvae to visualize the AKI mediated damage. https://www.leica-microsystems.com//science-lab/intravenous-microinjections-of-zebrafish-larvae-to-study-acute-kidney-injury/ Fri, 19 Jun 2015 20:26:00 +0000 https://www.leica-microsystems.com/15930 EM Sample Preparation Electron Backscatter Diffraction technique (EBSD) is known as a "surface" technique because electron diffraction is generated within a few tens of nanometers of the sample surface. Therefore, the specimen surface should be exempt of any damages in order to produce EBSD patterns. Here, we present a successful and efficient EBSD sample polishing of two very challenging specimens prepared by broad ion beam milling. https://www.leica-microsystems.com//science-lab/high-quality-sample-preparation-for-ebsd-analysis-by-broad-ion-beam-milling/ Fri, 12 Jun 2015 20:30:00 +0000 PhD Laurie Palasse, PhD Wolfgang Grünewald https://www.leica-microsystems.com/15458 Confocal Microscopy Fluorescence Microscopy Resolution, high signal intensity and elevated signal to noise ratio (SNR) are key issues for biologists who aim at studying the localisation of biological structures at the cellular and subcellular levels using confocal microscopy. The resolution required to separate sub-cellular biological structures is often near to the resolving power of the microscope. https://www.leica-microsystems.com//science-lab/improving-axial-resolution-in-confocal-microscopy-with-new-high-refractive-index-mounting-media/ Thu, 11 Jun 2015 13:04:00 +0000 https://www.leica-microsystems.com/15509 CLEM We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer superior structural preservation, performing in-resin super-resolution, however, remains a challenge. https://www.leica-microsystems.com//science-lab/correlative-in-resin-super-resolution-and-electron-microscopy-using-standard-fluorescent-proteins/ Thu, 04 Jun 2015 10:28:00 +0000 https://www.leica-microsystems.com/15227 EM Sample Preparation Capacitance and pyroelectric current measurements are best suited to determine important dielectric properties of materials, e.g. the relative dielectric constant, εr, and the remnant electric polarisation, P, of materials, respectively. In this technical report, we demonstrate the successful use of the Leica EM TXP surface targeting system to prepare a small capacitor. https://www.leica-microsystems.com//science-lab/preparation-of-small-disk-shaped-samples-for-capacitance-and-pyroelectric-current-measurements/ Fri, 29 May 2015 14:11:00 +0000 MSc, PhD, Prof. Andrea Gauzzi https://www.leica-microsystems.com/13642 CLEM EM Sample Preparation The urge to go deeper into the microscopic world has led researchers to combine the versatility of the light microscopy (LM) with the resolution power of the electron microscope (EM) to produce Correlative Light and Electron Microscopy (CLEM). CLEM’s most significant and powerful characteristic for cell biology research is an ability to study the same cell using two different microscopy platforms. The Essential Knowledge Briefing describes the basics of CLEM and also reviews potential pitfalls and problems as well as tricks for solving them. https://www.leica-microsystems.com//science-lab/correlative-light-and-electron-microscopy-get-your-free-clem-e-book-for-download/ Thu, 28 May 2015 16:34:00 +0000 Dr. Paul Verkade https://www.leica-microsystems.com/15767 Digital Microscopy Quality Assurance Digital microscopy is one of the buzz words in microscopy – and there are a couple of facts that are useful to know. Georg Schlaffer, Product Manager with Leica Microsystems, has often been asked about digital microscopy by customers and colleagues alike. He has worked together with Scientific Writer Jim DeRose for comprehensive answers to the most important ones. https://www.leica-microsystems.com//science-lab/what-you-always-wanted-to-know-about-digital-microscopy-but-never-got-around-to-asking/ Wed, 27 May 2015 11:40:00 +0000 PhD James DeRose, Georg Schlaffer https://www.leica-microsystems.com/15762 Confocal Microscopy Neuroscience Adult organisms have to adapt to survive, and the same is true for their tissues. Rates and types of cell production must be rapidly and reversibly adjusted to meet tissue demands in response to both local and systemic challenges. Recent work reveals how stem cell (SC) populations meet these requirements by switching between functional states tuned to homoeostasis or regeneration. https://www.leica-microsystems.com//science-lab/switching-roles-the-functional-plasticity-of-adult-tissue-stem-cells/ Mon, 18 May 2015 12:56:00 +0000 https://www.leica-microsystems.com/14978 Super-Resolution Confocal Microscopy Hendra virus (HeV) is a pleomorphic virus belonging to the Paramyxovirus family. Our long-term aim is to understand the process of assembly of HeV virions. As a first step, we sought to determine the most appropriate cell culture system with which to study this process, and then to use this model to define the morphology of the virus and identify the site of assembly by imaging key virus encoded proteins in infected cells. https://www.leica-microsystems.com//science-lab/detailed-morphological-characterisation-of-hendra-virus-infection-of-different-cell-types-using-super-resolution-and-conventional-imaging/ Wed, 13 May 2015 20:09:00 +0000 https://www.leica-microsystems.com/15606 Light Sheet Microscopy Confocal Microscopy Optical imaging instrumentation can magnify tiny objects, zoom in on distant stars and reveal details that are invisible to the naked eye. But it notoriously suffers from an annoying problem: the limited depth of field. Our eye-lens (an optical imaging instrument) has the same trouble, but our brain smartly removes all not-in-focus information before the signal reaches conscious cognition. https://www.leica-microsystems.com//science-lab/confocal-and-digital-light-sheet-imaging/ Mon, 11 May 2015 07:15:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Petra Haas https://www.leica-microsystems.com/14796 Super-Resolution Cell-sized polyelectrolyte capsules functionalized with a redox-driven proton pump protein were assembled for the first time. The interaction of polyelectrolyte microcapsules, fabricated by electrostatic layer-by-layer assembly, with cytochrome c oxidase molecules was investigated. We found that the cytochrome c oxidase retained its functionality, that the functionalized microcapsules interacting with cytochrome c oxidase were permeable and that the permeability characteristics of the microcapsule shell depend on the shell components. https://www.leica-microsystems.com//science-lab/multilayered-polyelectrolyte-microcapsules-interaction-with-the-enzyme-cytochrome-c-oxidase/ Tue, 05 May 2015 17:31:00 +0000 https://www.leica-microsystems.com/5080 Laser Microdissection There are many different types of consumables for laser microdissection (LMD) systems. They cover a wide range of applications from basic to highly specialized, enabling scientists to choose their own individual configuration whatever their field of research. https://www.leica-microsystems.com//science-lab/application-specific-consumables-for-laser-microdissection/ Mon, 04 May 2015 11:47:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/15608 Surgical Microscopy In this video, Dr. Ike Ahmed demonstrates how optical innovations help to improve ophthalmic surgeries. In addition, he talks about his experience with 3D heads-up surgery, using a stereoscopic high-definition visualization system for cataract, glaucoma, and retinal surgeries. https://www.leica-microsystems.com//science-lab/video-how-optical-innovations-and-3d-heads-up-surgery-help-to-improve-ophthalmic-surgery/ Mon, 27 Apr 2015 15:39:00 +0000 Dr. Ike Ahmed https://www.leica-microsystems.com/14920 Multiphoton Microscopy CLEM Live-Cell Imaging Cancer research unsing multiphoton microscopy and 3D electron microscopy. Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient events at high resolution. Performing correlative microscopy in complex and bulky samples such as an entire living organism is a time-consuming and error-prone task. https://www.leica-microsystems.com//science-lab/correlating-intravital-multi-photon-microscopy-to-3d-electron-microscopy-of-invading-tumor-cells-using-anatomical-reference-points/ Fri, 24 Apr 2015 13:06:00 +0000 https://www.leica-microsystems.com/14702 Education Confocal Microscopy Beyond the confines of the school curriculum, the young talent promotion program Initiative Junge Forscherinnen und Forscher e.V. (Initiative for Young Researchers) from Würzburg, Bavaria, has set itself the task of fostering an enthusiasm for natural sciences and future technologies in young people. Christoph Stolzenberger is one of the IJF’s science presenters. In his Experimentarium and NanoShuttle, he and his team of postgraduates inspire young researchers’ interest in the wonderful world of microstructures. https://www.leica-microsystems.com//science-lab/venturing-into-uncharted-dimensions-the-fascination-of-future-technologies/ Fri, 24 Apr 2015 12:29:00 +0000 Dr. Christoph Stolzenberger, MA Natalie Hampl https://www.leica-microsystems.com/15511 Super-Resolution The challenge of increasing the spatial resolution of an optical microscope beyond the diffraction limit can be reduced to a spectroscopy task by proper manipulation of the molecular states. The nanoscale spatial distribution of the molecules inside the detection volume of a scanning microscope can be encoded within the fluorescence dynamics and decoded by resolving the signal into its dynamics components. https://www.leica-microsystems.com//science-lab/encoding-and-decoding-spatio-temporal-information-for-super-resolution-microscopy/ Fri, 17 Apr 2015 10:35:00 +0000 https://www.leica-microsystems.com/15295 Fluorescence Microscopy Immunofluorescence (IF) is a powerful method for visualizing intracellular processes, conditions and structures. IF preparations can be analyzed by various microscopy techniques (e.g. CLSM, Epifluorescence, TIRF, GSDIM), depending on the application or the researcher’s interest. Meanwhile, IF has become indispensable for a large number of research groups which have at least access to a simple fluorescence microscope. https://www.leica-microsystems.com//science-lab/how-to-prepare-your-specimen-for-immunofluorescence-microscopy/ Mon, 13 Apr 2015 14:22:00 +0000 Florian Hoff https://www.leica-microsystems.com/15430 Multiphoton Microscopy Confocal Microscopy Live-Cell Imaging THG microscopy as special variants of multiphoton microscopy. Third Harmonic Generation (THG) microscopy is a non-fluorescent multi-photon technique that combines the advantages of label-free imaging with restriction of signal generation to the focal spot of the scanning laser. It allows three-dimensional imaging of refraction index mismatches and of hemoglobin. https://www.leica-microsystems.com//science-lab/third-harmonic-generation-microscopy-label-free-3d-tissue-imaging-and-blood-flow-characterization/ Sat, 11 Apr 2015 17:34:00 +0000 Dr. Steffen Dietzel https://www.leica-microsystems.com/15424 Stereo Microscopy The chick embryo and its circulatory system enclosed by the eggshell has become an important model for biomedical research such as the control of angiogenesis, cancer therapy, and drug delivery test. In this study, a novel artificial eggshell with functionalized surface is proposed, which allows the total amount of oxygen to pass into the egg for the chick embryo culturing and has high observability and accessibility for embryo manipulation. https://www.leica-microsystems.com//science-lab/egg-in-cube-design-and-fabrication-of-a-novel-artificial-eggshell-with-functionalized-surface/ Tue, 07 Apr 2015 15:55:00 +0000 https://www.leica-microsystems.com/15406 Super-Resolution Live-Cell Imaging Confocal Microscopy Multiphoton Microscopy Fluorescence Microscopy Quantitative Imaging Only a few days to go before the start of Focus on Microscopy 2015 in Göttingen, Germany. This year’s FOM is being organized by Dr. Gertrude Bunt and Prof. Dr. Fred S. Wouters from the University Medical Center, Göttingen, in cooperation with Prof. Dr. G.J. (Fred) Brakenhoff, University of Amsterdam, The Netherlands. https://www.leica-microsystems.com//science-lab/interview-with-dr-gertrude-bunt-and-prof-fred-s-wouters-on-the-fom-2015/ Fri, 27 Mar 2015 11:36:00 +0000 Dr. Gertrude Bunt, Dr. Fred S. Wouters, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/15404 Super-Resolution For Professor Stefan Hell, who won the Nobel Prize for Chemistry in 2014 for the development of super-resolved fluorescence microscopy and the development of STED microscopy, the Focus on Microscopy conference has a very special significance. It was at the very first FOM 1998 in Sydney where Hell gave one of his first lectures on super-resolution, entitled "Super-resolution through 4Pi-confocal microscopy in cellular imaging". https://www.leica-microsystems.com//science-lab/four-questions-for-professor-stefan-hell-on-the-subject-of-fom-2015/ Fri, 27 Mar 2015 10:54:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/15347 Super-Resolution The well-known saying of "Seeing is believing" became even more apt in biology when stimulated emission depletion (STED) nanoscopy was introduced in 1994 by the Nobel laureate S. Hell and coworkers. This article gives an overview of the various cutting-edge implementations of STED nanoscopy and tries to shine a light into the future: imaging everything faster with unprecedented sensitivity and label-free. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-a-glimpse-into-the-future/ Mon, 23 Mar 2015 18:08:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/14824 Laser Microdissection As most tissue specimens contain a mixture of different cell types including epithelial cells, stromal cells and immune cells, selection of the cells of interest is of utmost importance for the accurate determination of gene/microRNA expression. Laser capture microdissection enables the researcher to obtain homogeneous ultrapure cell selections from heterogeneous starting material. https://www.leica-microsystems.com//science-lab/total-rna-isolation-after-laser-capture-microdissection-of-human-cervical-squamous-epithelial-cells-from-fresh-frozen-tissue/ Fri, 20 Mar 2015 17:24:00 +0000 https://www.leica-microsystems.com/15106 EM Sample Preparation Fluorescence Microscopy Many biological insights can be obtained by combining the power of Fluorescence Microscopy (FM) with that of Electron Microscopy (EM) to study the same sample – this is called Correlative Light and Electron Microscopy (CLEM). In FM, specific proteins can be labelled and identified, and their dynamics and interactions can be visualized in fixed or living cells. https://www.leica-microsystems.com//science-lab/cryo-clem-the-combination-of-cryo-fluorescence-microscopy-with-cryo-electron-microscopy/ Wed, 18 Mar 2015 08:17:00 +0000 PhD Martin Schorb, PhD John A. G. Briggs https://www.leica-microsystems.com/15287 Quality Assurance Metallography With inverted microscopes, you look at samples from below since their optics are placed under the sample, with upright microscopes you look at samples from above. Traditionally, inverted microscopes are used for life science research, because gravity makes samples sink to the bottom of a holder with aqueous solution and you don’t see a lot from above. https://www.leica-microsystems.com//science-lab/five-advantages-of-inverted-over-upright-microscopes-in-industrial-applications/ Mon, 16 Mar 2015 09:06:00 +0000 Dr. Kay Scheffler, Claudia Müller https://www.leica-microsystems.com/15237 Super-Resolution Live-Cell Imaging STED nanoscopy has evolved to a highly versatile tool for the observation of the living cell, more and more finding its way into state-of-the-art optical imaging facilities in biomedical research institutes. https://www.leica-microsystems.com//science-lab/pathways-to-optical-sted-microscopy/ Thu, 12 Mar 2015 15:49:00 +0000 https://www.leica-microsystems.com/13946 TIRF Microscopy Total Internal Reflection Fluorescence (TIRF) Microscopy is a technique that only illuminates dye molecules near a surface. In this video, the pioneer of TIRF Microscopy describes what this technique is used for, explains the principles of the evanescent wave, gives many examples of different microscope configurations used in TIRF, and shows how polarized light TIRF can be used to image membrane orientation. https://www.leica-microsystems.com//science-lab/video-talk-by-daniel-axelrod-total-internal-reflection-fluorescence-tirf-microscopy/ Mon, 09 Mar 2015 08:45:00 +0000 https://www.leica-microsystems.com/14828 Laser Microdissection CLEM Confocal Microscopy Live-Cell Imaging Correlative light electron microscopy (CLEM) combines the advantages of light and electron microscopy, thus making it possible to follow dynamic events in living cells at nanometre resolution. Various CLEM approaches and devices have been developed, each of which has its own advantages and technical challenges. We here describe our customized patterned glass substrates, which improve the feasibility of correlative fluorescence/confocal and scanning electron microscopy. https://www.leica-microsystems.com//science-lab/customized-patterned-substrates-for-highly-versatile-correlative-light-scanning-electron-microscopy/ Fri, 06 Mar 2015 17:14:00 +0000 https://www.leica-microsystems.com/15057 Super-Resolution Live-Cell Imaging A SPAD array camera with single-photon sensitivity and zero read-out noise allows for the detection of extremely weak signals at ultra-fast imaging speeds. With temporal resolution in the order of micro-seconds, a SPAD array camera offers great potential for live-cell imaging with super-resolution. https://www.leica-microsystems.com//science-lab/towards-digital-photon-counting-cameras-for-single-molecule-optical-nanoscopy/ Wed, 04 Mar 2015 08:06:00 +0000 https://www.leica-microsystems.com/15111 Super-Resolution Super-resolution microscopy techniques have revolutionized biology for the last ten years. With their help cellular components can now be visualized at the size of a protein. Nevertheless, imaging living cells is a challenge for most of the super-resolution principles. https://www.leica-microsystems.com//science-lab/universal-paint-dynamic-super-resolution-microscopy/ Mon, 02 Mar 2015 08:24:00 +0000 Ph.D. Eric Hosy, Dr. Christoph Greb https://www.leica-microsystems.com/15132 Surface Metrology The excellent mechanical and electrical properties of carbon nanotubes have led to them being exploited for the creation of a new class of high performance polymer composites. Due to important advances in the last few years, nanotube containing polymers have been developed for optoelectronic applications. https://www.leica-microsystems.com//science-lab/nanoscale-or-microscale-structures-formed-in-polymers-containing-nanotubes-greatly-enhance-the-electrical-conductivity-potential-applications-for-photovoltaic-devices/ Fri, 27 Feb 2015 11:34:00 +0000 PhD James DeRose, Nicolas Boulanger, Patrice Belin, PhD David R. Barbero https://www.leica-microsystems.com/15203 Confocal Microscopy Live-Cell Imaging High speed scanning is necessary to image rapidly changing biological processes. With traditional scanning techniques, imaging speed is limited by the number of fluorophores in a specimen. And, rapid acquisition often comes at the cost of image resolution. https://www.leica-microsystems.com//science-lab/video-high-speed-scanning-with-two-scanners-in-one-system/ Tue, 24 Feb 2015 15:15:00 +0000 PhD Christopher Vega https://www.leica-microsystems.com/15081 EM Sample Preparation CLEM Cilia and flagella were the first organelles to be discovered and have been studied for centuries. However, their essential role in humans and how ciliary defects cause diseases are still not well understood. Cryo-EM has recently shed new light on their inner workings and solved some long-standing mysteries, only to raise new questions on how cilia and flagella function. https://www.leica-microsystems.com//science-lab/new-insights-into-cilia-and-flagella-by-cryo-em/ Mon, 16 Feb 2015 09:25:00 +0000 PhD Thomas Heuser https://www.leica-microsystems.com/12945 Super-Resolution GSDIM microscopy is a widefield super-resolution technique based on the localization of fluorophores with nanometer precision. With its help a lateral resolution of down to 20 nm can be achieved, whereas the new 3D feature even shrinks axial resolution to 50 nm. Here we provide a collection of publications around that super-resolution microscopy method also called dSTORM. https://www.leica-microsystems.com//science-lab/gsdim-publication-list/ Sun, 15 Feb 2015 10:25:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/15223 Super-Resolution Neuroscience The glass-supported planar lipid bilayer system has been utilized in a variety of disciplines. One of the most useful applications of this technique has been in the study of immunological synapse formation, due to the ability of the glass-supported planar lipid bilayers to mimic the surface of a target cell while forming a horizontal interface. https://www.leica-microsystems.com//science-lab/super-resolution-imaging-of-the-natural-killer-cell-immunological-synapse-on-a-glass-supported-planar-lipid-bilayer/ Thu, 12 Feb 2015 09:26:00 +0000 https://www.leica-microsystems.com/15104 EM Sample Preparation A well established usage case for cryo-TEM is three-dimensional reconstruction of isolated macromolecules, virus particles, or filaments. On one hand, these approaches are based on averaging of repetitive structures – either due to numerous identical molecules, repetitive patterns on a filament, or symmetries, to reduce the noise inherent to cryo-TEM. https://www.leica-microsystems.com//science-lab/immersion-freezing-for-cryo-transmission-electron-microscopy-applications/ Mon, 09 Feb 2015 14:51:00 +0000 Dr. Guenter Resch https://www.leica-microsystems.com/15102 EM Sample Preparation The high vacuum required in a transmission electron microscope (TEM) greatly impairs the ability to study specimens naturally occurring in an aqueous phase: exposing "wet" specimens to a pressure significantly lower than the vapour pressure of water will lead to the water phase boiling off rapidly in the column, with devastating consequences for the structure of the specimen. Hence, various methods to dry specimens before inspection are employed in conventional TEM – a preparative step often associated with artifacts limiting the significance of the results. https://www.leica-microsystems.com//science-lab/immersion-freezing-for-cryo-transmission-electron-microscopy-fundamentals/ Mon, 09 Feb 2015 14:41:00 +0000 Dr. Guenter Resch https://www.leica-microsystems.com/15154 Surgical Microscopy The lateral window technique is a method of maxillary sinus floor elevation which enables implant placement in the maxillary posterior region where the maxillary sinus is enlarged and/or the alveolar bone height is deficient. When creating a lateral window, great care should be taken not to perforate the maxillary sinus membrane. https://www.leica-microsystems.com//science-lab/video-implant-microsurgery-microscope-aided-maxillary-sinus-floor-elevation-lateral-window-technique/ Tue, 03 Feb 2015 09:09:00 +0000 DDS Eiju Sen https://www.leica-microsystems.com/15113 Live-Cell Imaging Fluorescence Microscopy Fighting cancer is a major goal of present-day medicine. So far mainly surgery, chemotherapy or radiation therapy are utilized to extinguish cancerous tissue, or at least set limits to it. Interestingly the human immune system has effective potential to fight cancer cells. Typically it reacts on parasitic, viral or bacterial infections. Thereby T-cells help to destroy infested cells after binding them via their specific antigen receptor. https://www.leica-microsystems.com//science-lab/immunotherapy-to-combat-cancer-sleeping-beauty-dna-plasmid-based-gene-transfer-system-to-modify-t-cells/ Wed, 28 Jan 2015 19:55:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/14890 EM Sample Preparation Confocal Microscopy The development of recombinant DNA technology has allowed the use of plants for the production of biopharmaceuticals. In contrast to other production platforms, plants are unexpensive, easy to scale up and lack human pathogens. Moreover, because plants are eukaryotes they can process and modify complex human proteins. https://www.leica-microsystems.com//science-lab/influence-of-tissue-and-plant-species-in-the-trafficking-of-a-recombinant-protein-in-plant-cells/ Mon, 26 Jan 2015 17:57:00 +0000 PhD Elsa Arcalis, PhD, MSc Eva Stöger https://www.leica-microsystems.com/14980 Super-Resolution Time-gated detection, namely, only collecting the fluorescence photons after a time-delay from the excitation events, reduces complexity, cost, and illumination intensity of a stimulated emission depletion (STED) microscope. In the gated continuous-wave- (CW-) STED implementation, the spatial resolution improves with increased time-delay, but the signal-to-noise ratio (SNR) reduces. https://www.leica-microsystems.com//science-lab/multi-images-deconvolution-improves-signal-to-noise-ratio-on-gated-stimulated-emission-depletion-microscopy/ Fri, 23 Jan 2015 12:22:00 +0000 https://www.leica-microsystems.com/15079 Super-Resolution Confocal Microscopy Since super-resolution has become one of the most favored methods in biomedical research, the term has become increasingly popular. Still, there is much of confusion about what is super-resolution and what is resolution at all. Here, the classical view of microscopic resolution is discussed and some techniques that resolve better than classical are briefly introduced. The picture on the right shows the intensity distribution of an image of two points whose distance is just the Rayleigh criterion (false color coding). https://www.leica-microsystems.com//science-lab/super-resolution-on-a-heuristic-point-of-view-about-the-resolution-of-a-light-microscope/ Wed, 21 Jan 2015 16:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/14918 Super-Resolution Live-Cell Imaging The aim of cell biology is to study smallest details on a cellular level preferably in a live cell experiment. By providing fast and direct super-resolution, STED (Stimulated Emission Depletion) microscopy is the perfect tool for studying cellular details in the nanometer range in vivo. https://www.leica-microsystems.com//science-lab/a-bright-dye-for-live-cell-sted-microscopy/ Mon, 19 Jan 2015 15:47:00 +0000 PhD Stefan Pitsch, PhD Isabelle Köster https://www.leica-microsystems.com/15077 Light Sheet Microscopy Just about everyone who has examined fluorescent samples under the microscope is aware of the constant struggle to have enough signal to see the labeled structures while also avoiding fluorophore bleaching. What may be less apparent, at least to those who image bright, robust or fixed samples, is how stressful and potentially toxic to living cells and tissues it is to illuminate them with high-intensity light. https://www.leica-microsystems.com//science-lab/nature-methods-light-sheet-fluorescence-microscopy-method-of-the-year-2014/ Wed, 14 Jan 2015 17:24:00 +0000 https://www.leica-microsystems.com/15059 Super-Resolution Diffraction plays a crucial role in microscopy as it prevents the recording of arbitrarily sharp images with conventional light microscopes. Many names are connected with the notion of diffraction and the definition of resolution. An overview over the contributions of the different scientists to the recognition and definition of the diffraction barrier in the past centuries is given and the recent developments that led to breaking this barrier are portrayed. https://www.leica-microsystems.com//science-lab/finding-defining-and-breaking-the-diffraction-barrier-in-microscopy-a-historical-perspective/ Mon, 12 Jan 2015 15:42:00 +0000 https://www.leica-microsystems.com/15219 Confocal Microscopy Audrey Salles is a specialist for confocal and super-resolution microscopy at Pasteur Institute, Imagopole, PFID, Paris, France. Her research interests are cytokine signaling and skeleton organization of human TCD4-cells. https://www.leica-microsystems.com//science-lab/leica-is-always-flexible-and-dynamic-interview-with-audrey-salles-pasteur-institute-paris/ Mon, 12 Jan 2015 11:10:00 +0000 Audrey Salles, PhD Isabelle Köster, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/13927 Basics in Microscopy An infinitesimally small point appears in the microscope as a spot with a certain size, blurred in the z-direction and with concentric rings around it. This "point spread function" reveals many of the optical properties of your microscope. This lecture explains why and how the microscope images a point as a point spread function. https://www.leica-microsystems.com//science-lab/video-talk-by-jeff-lichtman-point-spread-function/ Thu, 08 Jan 2015 13:23:00 +0000 Dr. Jeff Lichtman https://www.leica-microsystems.com/14858 Fluorescence Microscopy Confocal Microscopy In recent years, increasing numbers of human campylobacteriosis cases caused by contaminated water have been reported. As the culture-based detection of Campylobacter is time consuming and can yield false-negative results, the suitability of a quantitative real-time PCR method in combination with an ethidium monoazide pretreatment of samples (EMA-qPCR) for the rapid, quantitative detection of viable Campylobacter cells from water samples was investigated. https://www.leica-microsystems.com//science-lab/influencing-factors-and-applicability-of-the-viability-ema-qpcr-for-a-detection-and-quantification-of-campylobacter-cells-from-water-samples/ Tue, 06 Jan 2015 11:12:00 +0000 https://www.leica-microsystems.com/14367 Neuroscience Live-Cell Imaging Fluorescence Microscopy Confocal Microscopy Neural regeneration is a fascinating process with profound impact on human health, such that defining biological and genetic pathways is of interest. Here we describe an in vivo preparation for neuronal regeneration in the adult Drosophila. The nerve along the anterior margin of the wing is comprised of ~225 neurons that send projections into the central neuropil (thorax). https://www.leica-microsystems.com//science-lab/axon-injury-and-regeneration-in-the-adult-drosophila/ Fri, 02 Jan 2015 21:01:00 +0000 https://www.leica-microsystems.com/14955 Super-Resolution Jean-Luc Vonesch is head of the imaging facility at the Institute of Genetics and Molecular and Cellular Biology (IGBMC), Strasburg, France. 23 years ago he was the founder of this facility which nowadays serves more than 850 scientists distributed among 47 working groups. Looking deeply into the cells is of a special interest Vonesch states. And with super-resolution microscopy he pretends it is easier to identify the regions of interest for subsequent electron microscopy: “And so we can gain time thanks to the super-resolution” he says. https://www.leica-microsystems.com//science-lab/video-interview-with-jean-luc-vonesch/ Wed, 31 Dec 2014 11:27:00 +0000 Dr. Jean-Luc Vonesch, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/14774 Laser Microdissection Neuroscience Gliomas are mostly incurable secondary to their diffuse infiltrative nature. Thus, specific therapeutic targeting of invasive glioma cells is an attractive concept. As cells exit the tumor mass and infiltrate brain parenchyma, they closely interact with a changing micro-environmental landscape that sustains tumor cell invasion. https://www.leica-microsystems.com//science-lab/identification-of-molecular-pathways-facilitating-glioma-cell-invasion-in-situ/ Mon, 22 Dec 2014 09:35:00 +0000 https://www.leica-microsystems.com/14217 Super-Resolution Neuroscience In progressive multifocal leukoencephalopathy, JC virus–infected oligodendroglia display 2 distinct patterns of intranuclear viral inclusions: full inclusions in which progeny virions are present throughout enlarged nuclei and dot-shaped inclusions in which virions are clustered in subnuclear domains termed “promyelocytic leukemia nuclear bodies” (PML-NBs). Promyelocytic leukemia nuclear bodies may serve a scaffolding role in viral progeny production. https://www.leica-microsystems.com//science-lab/jc-virus-inclusions-in-progressive-multifocal-leukoencephalopathy-scaffolding-promyelocytic-leukemia-nuclear-bodies-grow-with-cell-cycle-transition-through-an-s-to-g2-like-state-in-enlarging-oligodendrocyte-nuclei/ Thu, 18 Dec 2014 13:31:00 +0000 https://www.leica-microsystems.com/14863 EM Sample Preparation Seeds provide a useful and versatile platform for the production of recombinant proteins and their numerous advantages have been often discussed. Among seeds, cereal crops offer additional advantages such as high yield and well-stablished agricultural infrastructure which allows easy up and down scaling in response to demand. https://www.leica-microsystems.com//science-lab/recovery-of-recombinant-antibody-is-affected-by-endogenous-protein-interaction-in-maize/ Mon, 15 Dec 2014 13:39:00 +0000 Dr. Jenny Peters https://www.leica-microsystems.com/14798 Quality Assurance Devices, products, and their components fabricated in many industries can be quite sensitive to contamination and, as a result, have stringent requirements for cleanliness. Measurement systems for automated particle analysis are often exploited for quantitative validation of product and component cleanliness to fulfill the needs of such industries as automotive, aerospace, microelectronics, pharmaceuticals, and medical devices. This report discusses the use of microscopy based measurement systems for automated particle analysis. https://www.leica-microsystems.com//science-lab/cleanliness-analysis-in-relation-to-particulate-contamination/ Mon, 08 Dec 2014 16:12:00 +0000 Yvonne Holzapfel, PhD James DeRose, Guido Kreck, PhD Markus Rochowicz https://www.leica-microsystems.com/14887 EM Sample Preparation High pressure freezing (HPF) is a cryo-fixation method primarily for biological samples, but also for a variety of non-biological materials. It is a technique that yields optimal preservation in many cell types and tissues or in organic and inorganic composites. Most commonly, the high pressure frozen samples are analyzed further with light or electron microscopy after appropriate processing. https://www.leica-microsystems.com//science-lab/video-tutorials-filling-and-assembling-of-different-carriers-for-high-pressure-freezing/ Fri, 05 Dec 2014 08:45:00 +0000 Dr. Cveta Tomova https://www.leica-microsystems.com/14884 Confocal Microscopy Super-Resolution Live-Cell Imaging Multiphoton Microscopy Fluorescence Microscopy Quantitative Imaging Neuroscience Confocal microscopy has come a very long way since its invention more than a half-century ago. Today, with novel technology driven by leading imaging companies, it has become the standard for fluorescence microscopy. Choosing the right confocal microscope for your specific research requires the appropriate mix of features related to resolution, sensitivity, and speed. https://www.leica-microsystems.com//science-lab/how-to-choose-the-right-confocal-microscope-for-your-lab/ Wed, 03 Dec 2014 15:15:00 +0000 PhD Christopher Vega https://www.leica-microsystems.com/13911 Super-Resolution Live-Cell Imaging We describe for the first time the combination between cross-pair correlation function analysis (pair correlation analysis or pCF) and stimulated emission depletion (STED) to obtain diffusion maps at spatial resolution below the optical diffraction limit (super-resolution). Our approach was tested in systems characterized by high and low signal to noise ratio, i.e. Capsid Like Particles (CLPs) bearing several (>100) active fluorescent proteins and monomeric fluorescent proteins transiently expressed in living Chinese Hamster Ovary cells, respectively. https://www.leica-microsystems.com//science-lab/nanoscale-protein-diffusion-by-sted-based-pair-correlation-analysis/ Fri, 21 Nov 2014 09:36:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/14451 Super-Resolution Until recently, the diffraction of light had placed a fundamental limit on how far biologists could peer into cells with optical microscopes, preventing them from resolving features less than 250 nm in size, missing critical structures within cells. Over the past 20 years scientists have developed several ingenious techniques allowing them to resolve features as small as 20 nm. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-get-your-free-e-book-for-download/ Wed, 19 Nov 2014 15:10:00 +0000 https://www.leica-microsystems.com/13948 Neuroscience Optogenetics is a combination of genetics and optics to achieve a gain or loss of function of biochemical events such as action potentials in a particular neuron or tissue. Opsin genes encode proteins that receive light and give rise to ion flow. This talk gives an introduction to optogenetics followed by examples of how optogenetics is being used to study the brain. https://www.leica-microsystems.com//science-lab/video-talk-by-karl-deisseroth-optogenetics/ Tue, 18 Nov 2014 12:52:00 +0000 https://www.leica-microsystems.com/14455 EM Sample Preparation Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D) organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. https://www.leica-microsystems.com//science-lab/electron-tomography-of-cryo-immobilized-plant-tissue-a-novel-approach-to-studying-3d-macromolecular-architecture-of-mature-plant-cell-walls-in-situ/ Mon, 17 Nov 2014 17:47:00 +0000 https://www.leica-microsystems.com/14727 Confocal Microscopy Multiphoton Microscopy CLARITY clearing derivatives for multiphoton microscopy. Transparent organisms help us to identify spatial arrangements and connections of cells and tissues, especially neuronal circuits can easily be identified and characterized. CLARITY is on everyone's lips. https://www.leica-microsystems.com//science-lab/deeper-insights-in-transparent-animals/ Mon, 10 Nov 2014 09:48:00 +0000 BS, PhD Viviana Gradinaru, PhD Isabelle Köster https://www.leica-microsystems.com/14656 Confocal Microscopy Super-Resolution CLEM Live-Cell Imaging Quantitative Imaging To optimise the efficiency of cell machinery, cells can use the same protein (often called a hub protein) to participate in different cell functions by simply changing its target molecules. There are large data sets describing protein-protein interactions ("interactome") but they frequently fail to consider the functional significance of the interactions themselves. https://www.leica-microsystems.com//science-lab/icln-a-new-regulator-of-non-erythroid-41r-localisation-and-function/ Mon, 03 Nov 2014 18:02:00 +0000 https://www.leica-microsystems.com/14693 Stereo Microscopy Jenny Natusch from Lancaster, UK, is an artist, a photographer, a microphotographer and most of all she is a "sandgazer". With her totally different way of looking at things, she creates wonderful detailed images and photographs out of the dusty, brown mass we call sand, using a microscope to uncover it’s hidden beauty from all over the world. https://www.leica-microsystems.com//science-lab/welcome-to-sandland-how-microscopy-uncovers-the-beauty-of-sand/ Thu, 30 Oct 2014 12:35:00 +0000 Jenny Natusch, MA Natalie Hampl https://www.leica-microsystems.com/13917 Confocal Microscopy Live-Cell Imaging Diagnosing bacterial infection (BI) remains a challenge for the attending physician. An ex vivo infection model based on human fixed polymorphonuclear neutrophils (PMNs) gives an autofluorescence signal that differs significantly between stimulated and unstimulated cells. We took advantage of this property for use in an in vivo pneumonia mouse model and in patients hospitalized with bacterial pneumonia. https://www.leica-microsystems.com//science-lab/analysis-of-autofluorescence-in-polymorphonuclear-neutrophils-a-new-tool-for-early-infection-diagnosis/ Wed, 29 Oct 2014 12:21:00 +0000 https://www.leica-microsystems.com/14662 Basics in Microscopy Polarized light microscopy (PLM) has often been utilized in the fields of geology and the material sciences with significant implications in determining the mineral compositions and structures of composite materials (e.g. fibers) through the use of typical quartz wedge compensators, mica quarter-waveplates and de Sénarmont compensation. https://www.leica-microsystems.com//science-lab/micropolarimetric-analysis-of-guard-cell-walls-in-m-koenigii-versus-de-novo-localized-starch-granules-in-freshly-isolated-potato-tuber/ Wed, 22 Oct 2014 13:39:00 +0000 Shiraz S. Kaderuppan https://www.leica-microsystems.com/14614 Super-Resolution In this list you can find a compilation of scientific publications, interviews and videos related to Stefan Hella s well as key publications of the Nobel Prize and selected papers on STED and GSDIM super-resolution microscopy. https://www.leica-microsystems.com//science-lab/papers-and-articles-on-the-nobel-prize-in-chemistry-2014-stefan-hell-and-super-resolution-microscopy/ Fri, 17 Oct 2014 14:26:00 +0000 https://www.leica-microsystems.com/14658 Super-Resolution Neuroscience The molecular composition of the organelles involved in membrane recycling is difficult to establish as a result of the absence of suitable labeling tools. We introduce in this paper a novel probe, named membrane-binding fluorophore-cysteine-lysine-palmitoyl group (mCLING), which labels the plasma membrane and is taken up during endocytosis. https://www.leica-microsystems.com//science-lab/a-new-probe-for-super-resolution-imaging-of-membranes-elucidates-trafficking-pathways/ Fri, 17 Oct 2014 12:16:00 +0000 https://www.leica-microsystems.com/14386 EM Sample Preparation Coating of samples is required in the field of electron microscopy to enable or improve the imaging of samples. Compared to the traditional coater design, all parts of a Leica EM ACE Coater can be individually removed and cleaned or, if special cleanliness is needed, even exchanged for spare parts. https://www.leica-microsystems.com//science-lab/how-to-clean-a-coater/ Thu, 16 Oct 2014 16:24:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/14647 Super-Resolution The resolution needed to image subcellular architecture and dynamics in light microscopy is hindered by the diffraction limits as described by Ernst Abbe. Simply stated, structures smaller than 200 nanometers are lost in a blur. However, the field of super-resolution microscopy has produced methods to obtain resolution beyond this limit. Leica Microsystems has pioneered this field and offers the Leica TCS SP8 STED 3X for 3D Stimulated Emission Depletion microscopy. STED instantly produces super-resolution images, compatible with the dynamics of living cells, without the need for post-processing. https://www.leica-microsystems.com//science-lab/3d-sted-stimulated-emission-depletion-microscopy/ Tue, 14 Oct 2014 14:03:00 +0000 PhD Christopher Vega https://www.leica-microsystems.com/14623 Super-Resolution On October 8th 2014, The Royal Swedish Academy of Sciences has decided to award the Nobel Prize in Chemistry for 2014 to Eric Betzig, Stefan W. Hell and William E. Moerner "for the development of super-resolved fluorescence microscopy". For a long time optical microscopy was held back by a presumed limitation: that it would never obtain a better resolution than half the wavelength of light. Helped by fluorescent molecules the Nobel Laureates in Chemistry 2014 ingeniously circumvented this limitation. Their ground-breaking work has brought optical microscopy into the nanodimension. https://www.leica-microsystems.com//science-lab/nobel-prize-in-chemistry-for-achievements-in-super-resolution-microscopy/ Thu, 09 Oct 2014 13:25:00 +0000 Nobel Media AB 2014 https://www.leica-microsystems.com/14612 Surgical Microscopy Neurosurgeons and ophthalmologists deal with delicate structures, deep or narow cavities and tiny structures with vitally important functions. A clear, three-dimensional view on the surgical field is thus indispensable for the outcome of the operation and the patient’s safety. Until now, an increased depth of field for a larger three-dimensional area in focus was only achievable by reducing the resolution. A new technology is able to overcome this challenge. https://www.leica-microsystems.com//science-lab/fusionoptics-in-neurosurgery-and-ophthalmology-for-a-larger-3d-area-in-focus/ Wed, 08 Oct 2014 19:17:00 +0000 https://www.leica-microsystems.com/13795 Super-Resolution Professor Stefan Hell is director at the Max Planck Institute for Biophysical Chemistry and head of the department of NanoBiophotonics in Goettingen and widely considered as the father of super-resolution. His inventions of 4Pi and STED microscopy were turned into the first commercial super-resolution microscopes available by Leica Microsystems in 2004 and 2007. https://www.leica-microsystems.com//science-lab/video-interview-with-stefan-hell-the-inventor-of-super-resolution/ Wed, 08 Oct 2014 18:14:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell, Dipl. oec.-troph. Anja Schué, PhD Isabelle Köster https://www.leica-microsystems.com/14594 Surgical Microscopy Ergonomics Pain and muscle tension are common phenomena in a neurosurgeon’s workday: 78% of neurosurgeons surveyed reported pain after a day of surgery, 83% of these had musculoskeletal pain. By its nature, neurosurgery requires compromising positions. However, with a few simple steps and the right ergonomic accessories for the surgical microscope, the working environment in the OR can be made as comfortable as possible. https://www.leica-microsystems.com//science-lab/7-tips-for-better-ergonomics-in-neurosurgery/ Wed, 08 Oct 2014 17:37:00 +0000 https://www.leica-microsystems.com/14406 EM Sample Preparation Freeze fracture describes the technique of breaking a frozen specimen to reveal internal structures. Freeze etching is the sublimation of surface ice under vacuum to reveal details of the fractured face that were originally hidden. A metal/carbon mix enables the sample to be imaged in a SEM (block-face) or TEM (replica). It is used to investigate for instance cell organelles, membranes, layers and emulsions. https://www.leica-microsystems.com//science-lab/brief-introduction-to-freeze-fracture-and-etching/ Wed, 01 Oct 2014 09:27:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/14492 Fluorescence Microscopy Scientists at the University of Göttingen have deciphered the biogenesis of an enzyme complex whose role is to ensure that the ends of chromosomes are not shortened during the cell division process and that the genetic material is fully maintained. The results were published in Cell Reports magazine. https://www.leica-microsystems.com//science-lab/defect-transport-mechanism-leads-to-shortened-chromosome-ends/ Fri, 26 Sep 2014 16:46:00 +0000 Haijia Wu https://www.leica-microsystems.com/14389 Super-Resolution TIRF Microscopy William Hughes works at the Garvan Institute of Medical Research, Sydney (Australia). In his Lab Head position he is interested in the causes of diabetes particularly looking at changes in exocytic behavior of pancreatic beta cells as well as fat and muscle cells. TIRF microscopy is predestined for researchers looking at cellular processes near the cytoplasmic membrane. https://www.leica-microsystems.com//science-lab/video-interview-with-william-hughes/ Wed, 24 Sep 2014 15:14:00 +0000 Dr. William Hughes, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/14347 EM Sample Preparation Before ultrathin sectioning a sample with an ultramicrotome it has to be pre-prepared. For this pre-preparation, special attention must be paid to the sample size (size of the section), location of the sample (targeting) and accuracy of the block-face edges. This process is generally called trimming, wherein the sample is shaped mainly to a frustum of a pyramid. https://www.leica-microsystems.com//science-lab/brief-introduction-to-specimen-trimming/ Mon, 22 Sep 2014 13:56:00 +0000 Robert Ranner https://www.leica-microsystems.com/14402 Multiphoton Microscopy Neuroscience Multi-channel multiphoton microscopy with dedicated optics for CLARITY. Why clearing? Curiosity is human nature. And nothing attracts as much curiosity as the inside of living organisms. While in ancient times those who cut human bodies open to do research were put to death, and modern anatomy started only after Pope Clement VII allowed dissection, we can now watch brains working in living animals – and have a good chance of soon being able to interfere with the observed activities for healing (or control) purposes. https://www.leica-microsystems.com//science-lab/clearing-procedures-for-deep-tissue-imaging/ Thu, 18 Sep 2014 12:36:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Andrea Mülter https://www.leica-microsystems.com/13883 Super-Resolution The first super-resolution image he saw was an eye opener for him: "It was not just structures that got smaller. I was looking at a sample that I specifically had high hopes of seeing another layer of complexity and this actually was there." https://www.leica-microsystems.com//science-lab/video-interview-with-timo-zimmermann/ Wed, 10 Sep 2014 09:19:00 +0000 Dr. Timo Zimmermann, Dipl. oec.-troph. Anja Schué, PhD Isabelle Köster https://www.leica-microsystems.com/15992 Digital Microscopy Quality Assurance For certain applications, such as the production and maintenance of automobiles, aircraft, or power plants, quality control and reliability assurance, as well as safety and health inspection, accurate and reliable image data with precise calibration are very important. Digital microscopes are appealing for a wide range of technical applications in various industries, such as automotive, aerospace, precision engineering, microelectronics, and medical devices. https://www.leica-microsystems.com//science-lab/are-your-digital-microscope-measurements-accurate-and-reliable/ Mon, 08 Sep 2014 11:01:00 +0000 PhD James DeRose, Georg Schlaffer https://www.leica-microsystems.com/14343 EM Sample Preparation Glycerol spraying/low angle rotary shadowing (Aebi and Baschong, 2006) is a preparation technique used in biology to visualize structures yielding insufficent contrast with other techniques, due to their small diameter. This method is commonly used for specimens which include proteins with coiled coil domains or DNA. https://www.leica-microsystems.com//science-lab/glycerol-sprayingplatinum-low-angle-rotary-shadowing-of-dna-with-the-leica-em-ace600-e-beam/ Thu, 04 Sep 2014 14:33:00 +0000 Dr. Guenter Resch https://www.leica-microsystems.com/13879 Super-Resolution Kees Jalink's group at the Netherlands Cancer Institute in Amsterdam, The Netherlands, explores signal transduction pathways and cell adhesion processes in cancer cells. In his eyes especially the new three-dimensional nanoscopic view of the relevant structure of interest is an essential feature to get the full picture. https://www.leica-microsystems.com//science-lab/video-interviews-with-kees-jalink/ Fri, 29 Aug 2014 11:58:00 +0000 PhD Kees Jalink, MSc Leila Nahidiazar, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/14080 Fluorescence Microscopy Live-Cell Imaging The cephalochordate Amphioxus naturally co-expresses fluorescent proteins (FPs) with different brightness, which thus offers the rare opportunity to identify FP molecular feature/s that are associated with greater/lower intensity of fluorescence. Here, we describe the spectral and structural characteristics of green FP (bfloGFPa1) with perfect (100%) quantum efficiency yielding to unprecedentedly-high brightness, and compare them to those of co-expressed bfloGFPc1 showing extremely-dim brightness due to low (0.1%) quantum efficiency. https://www.leica-microsystems.com//science-lab/spectral-and-structural-comparison-between-bright-and-dim-green-fluorescent-proteins-in-amphioxus/ Thu, 28 Aug 2014 15:54:00 +0000 https://www.leica-microsystems.com/14246 Super-Resolution Neuroscience The isotopic composition of different materials can be imaged by secondary ion mass spectrometry. In biology, this method is mainly used to study cellular metabolism and turnover, by pulsing the cells with marker molecules such as amino acids labelled with stable isotopes (15N, 13C). The incorporation of the markers is then imaged with a lateral resolution that can surpass 100 nm. https://www.leica-microsystems.com//science-lab/correlated-optical-and-isotopic-nanoscopy/ Fri, 22 Aug 2014 09:53:00 +0000 https://www.leica-microsystems.com/14215 Multiphoton Microscopy Neuroscience Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. https://www.leica-microsystems.com//science-lab/single-cell-phenotyping-within-transparent-intact-tissue-through-whole-body-clearing/ Wed, 20 Aug 2014 11:26:00 +0000 https://www.leica-microsystems.com/11763 Basics in Microscopy Light has properties of particles and waves. Understanding the wave nature of light is essential to understanding the workings of a microscope. This lecture describes Huygens Wavelets, constructive/destructive interference, and diffraction. https://www.leica-microsystems.com//science-lab/video-talk-by-jeff-lichtman-resolution-in-microscopy-wave-optics-and-the-diffraction-limit/ Thu, 14 Aug 2014 12:48:00 +0000 Dr. Jeff Lichtman https://www.leica-microsystems.com/13950 Super-Resolution Neuroscience Recent developments in fluorescence far-field microscopy such as STED microscopy have accomplished observation of the living cell with a spatial resolution far below the diffraction limit. Here, we briefly review the current approaches to super-resolution optical microscopy and present the implementation of STED microscopy for novel insights into live cell mechanisms, with a focus on neurobiology and plasma membrane dynamics. https://www.leica-microsystems.com//science-lab/sted-microscopy-of-living-cells-new-frontiers-in-membrane-and-neurobiology/ Tue, 12 Aug 2014 12:11:00 +0000 Prof. Christian Eggeling https://www.leica-microsystems.com/14156 Forensics To be able to optically compare two objects with scientific accuracy, it must be possible to view them at the same time. This is particularly true for comparing small objects that can only be visualized with the aid of an optical magnification system. If you only have one microscope and have to look at the objects alternately, you need an excellent memory and can never rule out the risk of judgment errors, especially when examining objects with only minor differences in structure, color or profile. https://www.leica-microsystems.com//science-lab/125-years-of-comparison-microscopy-from-the-camera-lucida-to-the-automated-comparison-bridge/ Tue, 12 Aug 2014 09:11:00 +0000 Rolf Beck https://www.leica-microsystems.com/14017 Super-Resolution The 4th Super-Resolution User Club Meeting was held in collaboration with Christian Eggeling and the Weatherall Institute of Molecular Medicine in Oxford, UK. Here we present the abstracts of the talks and interviews with participants. https://www.leica-microsystems.com//science-lab/abstracts-of-the-4th-european-super-resolution-user-club-meeting/ Fri, 08 Aug 2014 08:09:00 +0000 Prof. Christian Eggeling, Ph.D. Giuseppe Vicidomini, MSc Leila Nahidiazar, Dr. Sergi Padilla-Parra, Prof. Mark Neil, Dr. Marko Lampe, PhD Kees Jalink, Dr. Katrin Willig, Dr. Timo Zimmermann, Ph.D. Marc van Zandvoort https://www.leica-microsystems.com/14066 Multiphoton Microscopy Live-Cell Imaging A recent publication in Nature shows that all stem cells divide and compete for niche space by passively "kicking out" others so that eventually one stem cell takes over the whole niche. Jacco van Rheenen and Saskia Ellenbroek talk about a new method of intravital imaging, which allows following the fate of individual stem cells over time in vivo and explains the new paradigm for stem cell development in the intestinal stem cell niche. https://www.leica-microsystems.com//science-lab/the-environment-makes-the-stem-cell/ Mon, 04 Aug 2014 16:25:00 +0000 PhD Jacco van Rheenen, PhD Saskia I. J. Ellenbroek, PhD Isabelle Köster https://www.leica-microsystems.com/14063 Confocal Microscopy Multiphoton Microscopy High time-resolution confocal microscopy (HTRCLSM) requires fast scanning devices. Whereas non-resonant galvo scanners allow full position control, but only at slow speed, resonant scanners allow ~25,000 lines per second, but offer much less positioning freedom. To still allow zoom and pan functions, several approaches have been tried, with varying success. The Leica confocal microscopes od the TCS series use a very smart solution that enables stepless zooming with short switching times. https://www.leica-microsystems.com//science-lab/smart-control-for-resonant-galvo-scanners/ Fri, 01 Aug 2014 08:29:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/12777 Neuroscience This video features a few of the major scientists and findings that have contributed to modern neuroscience: The history of our knowledge of the brain, how technical and conceptual advances led to new insights and the human side of neuroscience through quotations from key individuals on the nature of the brain and consciousness. https://www.leica-microsystems.com//science-lab/video-the-ascent-a-brief-history-of-the-brain/ Thu, 31 Jul 2014 06:04:00 +0000 https://www.leica-microsystems.com/13782 Laser Microdissection In contrast to other systems which use a fixed laserfocus for dissection, Leica Microsystems' LMD systems guide the laserfocus for dissection. This unique feature allows highly precise laser dissection independent of the stage accuracy. This tutorial describes the basic principles of the laser guidance and how the laser can be adjusted for perfect cutting results. https://www.leica-microsystems.com//science-lab/the-basic-principles-of-laser-microdissection/ Thu, 24 Jul 2014 13:48:00 +0000 Dr. Falk Schlaudraff https://www.leica-microsystems.com/13913 Laser Microdissection This study was designed to investigate the activation of the unfolded protein response (UPR) in tumor associated endothelial cells (TECs) and its association with chemoresistance during acidic pH stress. https://www.leica-microsystems.com//science-lab/glucose-regulated-protein-78-grp78-confers-chemoresistance-to-tumor-endothelial-cells-under-acidic-stress/ Mon, 21 Jul 2014 17:16:00 +0000 https://www.leica-microsystems.com/13839 Super-Resolution Super-resolution (SR) fluorescence microscopy has been revolutionizing the way in which we investigate the structures, dynamics, and functions of a wide range of nanoscale systems. In this review, I describe the current state of various SR fluorescence microscopy techniques along with the latest developments of fluorophores and labeling for the SR microscopy. https://www.leica-microsystems.com//science-lab/super-resolution-molecular-and-functional-imaging-of-nanoscale-architectures-in-life-and-materials-science/ Mon, 07 Jul 2014 11:45:00 +0000 https://www.leica-microsystems.com/13877 Super-Resolution Neuroscience A research team from Göttingen, led by Prof. Silvio O. Rizzoli, managed to determine the copy numbers and positions of all important building blocks of a synapse for the first time. This allowed them to reconstruct the first scientifically accurate 3D model of a synapse. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-helped-to-create-the-first-3d-model-of-a-synapse/ Tue, 01 Jul 2014 14:59:00 +0000 https://www.leica-microsystems.com/13837 EM Sample Preparation At the Institut de Minéralogie, de Physique des Matériaux et de Cosmochimie (IMPMC) of the Université Pierre et Marie Curie in Paris an new lab for EM Sample Preparation welcomes researchers of materials sciences and mineralogy: the lab has been equipped with Leica EM TXP target surfacing systems and Leica EM TIC3X ion beam systems. https://www.leica-microsystems.com//science-lab/partner-lab-for-em-sample-preparation-in-paris/ Fri, 27 Jun 2014 07:13:00 +0000 Guillaume Fiquet https://www.leica-microsystems.com/13495 Laser Microdissection Secondary plant metabolites are involved in various plant physiological and biochemical processes, allowing plants to interact successfully with their biotic and abiotic environment. Distribution patterns of these compounds at the tissue and cellular level show how plant defense systems are organized to manage environmental challenges in a targeted and precise way. https://www.leica-microsystems.com//science-lab/preparation-of-laser-microdissected-plant-cell-and-tissue-material-for-microchemical-analysis/ Tue, 24 Jun 2014 09:08:00 +0000 PhD Dirk Hölscher, PhD Sara Agnolet, PhD Jingjing Fang, PhD Jonathan Gershenzon, PhD Oliver Kayser, PhD Sheng-Hong Li, PhD Bernd Schneider https://www.leica-microsystems.com/13800 EM Sample Preparation For some years now, the search for alternative sources of raw materials has concentrated on oil shale deposits. As the exploitation of these raw materials is still extremely complex and expensive, analysis of oil shale samples is therefore extremely important for identifying the deposit’s potential in advance and optimizing mining methods. https://www.leica-microsystems.com//science-lab/analysis-of-oil-shale-as-an-alternative-source-of-energy/ Tue, 24 Jun 2014 08:23:00 +0000 PhD Wolfgang Grünewald https://www.leica-microsystems.com/13786 EM Sample Preparation Metallography The microstructure of new materials with heterogenic components is extremely difficult to study. As mechanical polishing often not leads to smooth surfaces, a novel ion beam cutting technique has proven to be very effective. In this article, the applicability of a novel triple ion beam (TIB) cutting technique for a clean metallographic preparation of the diamond/Al composites will be described. https://www.leica-microsystems.com//science-lab/triple-ion-beam-cutting-of-diamondal-composites-for-interface-characterization/ Sat, 21 Jun 2014 09:22:00 +0000 PhD Gang Ji, PhD Wolfgang Grünewald https://www.leica-microsystems.com/13527 Super-Resolution Historically, light microscopy has been limited in its ability to resolve closely spaced objects, with the best microscopes only able to resolve objects separated by 200 nm or more. This limit is known as the diffraction limit. In the last twenty years, a number of techniques have been developed that allow resolution beyond the diffraction limit. https://www.leica-microsystems.com//science-lab/video-talk-on-super-resolution-overview-and-stimulated-emission-depletion-sted-microscopy/ Thu, 05 Jun 2014 17:05:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/13490 Surface Metrology Thin film characterization technologies are in high demand, given the wide-spread use of coatings in all engineering and science fields. The properties of thin films can vary dramatically, i.e. thickness, optical and electrical properties, hardness, etc., that is difficult to find a general purpose characterization technique. https://www.leica-microsystems.com//science-lab/characterization-of-thin-films-using-high-definition-confocal-microscopy/ Thu, 05 Jun 2014 10:52:00 +0000 MSc Marco Renzelli, PhD Edoardo Bemporad https://www.leica-microsystems.com/13740 Super-Resolution Neuroscience Yasushi Okada, team leader at Riken Quantitative Biology Center in Osaka, Japan, investigates vesicular transport mechanisms in neuronal cells. As the size of transported vesicles is below 100 nm and the diameter of microtubules is about 25 nm, he uses super-resolution techniques to study the sophisticated machinery of neuronal transport. https://www.leica-microsystems.com//science-lab/interview-with-dr-yasushi-okada/ Tue, 03 Jun 2014 15:38:00 +0000 Dipl. oec.-troph. Anja Schué, MD, PhD Yasushi Okada, PhD Isabelle Köster https://www.leica-microsystems.com/13373 Super-Resolution Neuroscience Synaptic vesicle recycling is one of the best‐studied cellular pathways. Many of the proteins involved are known, and their interactions are becoming increasingly clear. However, as for many other pathways, it is still difficult to understand synaptic vesicle recycling as a whole. https://www.leica-microsystems.com//science-lab/synaptic-vesicle-recycling-steps-and-principles/ Mon, 26 May 2014 09:30:00 +0000 Prof. Silvio Rizzoli https://www.leica-microsystems.com/13555 Education Basics in Microscopy Learning begins with perception. Sensory impressions are branded in our mind and become the building blocks of knowledge. The more intensively young people are involved in the lesson and the more experiences they can make themselves, the easier they find it to learn. Hands-on microscopy is therefore a key component of modern science teaching, for example at the Philippinum Grammar School in Weilburg, where students love working with Leica educational microscopes. https://www.leica-microsystems.com//science-lab/hunting-down-the-hay-bacillus-educational-microscopes-in-biology-teaching/ Tue, 20 May 2014 17:16:00 +0000 MA Natalie Hampl https://www.leica-microsystems.com/13557 Education Basics in Microscopy Stereo Microscopy The Vogel- und Naturschutz-Tierpark Herborn to the north of Frankfurt may be small, but it’s always a great experience for school classes – and not only because it’s home to more than 300 animals of 80 different species, from South African blue cranes and white storks to meerkats and muntjac deer. Another special highlight for schoolchildren is the opportunity to look through a stereomicroscope and gain fascinating insights into nature. https://www.leica-microsystems.com//science-lab/bird-park-gives-fascinating-insights-into-the-variety-of-nature/ Mon, 19 May 2014 09:43:00 +0000 M. A. Janika Wiesner, Britta Löbig https://www.leica-microsystems.com/13487 Surface Metrology Porcelain stoneware is a product for which the synergy between the manufacturing technology and physical-mechanical properties has been particularly well developed with excellent results. Despite its excellent technical characteristics, the slip resistance can be significantly different depending on the tile surface finish: structured or smooth, glazed or polished. https://www.leica-microsystems.com//science-lab/anti-slip-treatment-on-glazed-ceramic-tiles-microstructural-and-topographic-modifications/ Wed, 14 May 2014 13:26:00 +0000 PhD Elisa Rambaldi, Barbara Mazzanti https://www.leica-microsystems.com/13280 EM Sample Preparation Neuroscience The combination of two powerful techniques: optogenetics and high-pressure freezing now makes it possible to visualize a dynamic cellular activity with temporal resolution of 5 milliseconds. By coupling a flash of light with high-pressure freezing, the process of vesicle recycling at the synapses can now be imaged by electron microscopy. https://www.leica-microsystems.com//science-lab/capturing-cellular-dynamics-with-millisecond-temporal-resolution/ Mon, 12 May 2014 13:16:00 +0000 PhD Shigeki Watanabe, PhD Erik M. Jørgensen https://www.leica-microsystems.com/13439 Surgical Microscopy If medication and laser techniques have no prospect of success in the treatment of glaucoma, surgery is essential to stop further damage to the optic nerve. A new surgical method is showing promising results for reducing the potential complications of an operation. Professor Dr. Dr. h.c. Franz Grehn, former Director of Würzburg University Eye Hospital, has developed the new method and produced a video to accompany a clinical comparison study. This film was awarded the Video Award 2013 sponsored by Leica Microsystems at the 111th Congress of the German Ophthalmological Society. https://www.leica-microsystems.com//science-lab/new-paths-in-glaucoma-therapy-operation-video-supports-knowledge-transfer-in-eye-surgery/ Wed, 07 May 2014 08:31:00 +0000 PhD Franz Grehn https://www.leica-microsystems.com/13034 Confocal Microscopy The objective of this study was to establish a primary culture of omasal epithelial cells (OECs) derived from dairy calves and to characterize its function in small peptide absorption. Bovine omasal tissues were obtained from newborn Chinese Holstein calves and digested with a 2.5% trypsin solution to obtain OECs. https://www.leica-microsystems.com//science-lab/establishment-and-characterization-of-an-omasal-epithelial-cell-model-derived-from-dairy-calves-for-the-study-of-small-peptide-absorption/ Mon, 05 May 2014 16:46:00 +0000 https://www.leica-microsystems.com/12428 Confocal Microscopy Fluorescence Microscopy The ability to colonize the host plants’ rhizospheres is a crucial feature to studyin the case of Plant Growth Promoting Rhizobacteria (PGPRs) with potential agricultural applications. In this work, we have created GFP-tagged derivatives of three candidate PGPRs: Bacillus subtilis MB73/2, Pseudomonas sp. P482 and Ochrobactrum sp. A44. https://www.leica-microsystems.com//science-lab/colonization-of-potato-rhizosphere-by-gfp-tagged-bacillus-subtilis-mb732-pseudomonas-sp-p482-and-ochrobactrum-sp-a44-shown-on-large-sections-of-roots-using-enrichment-sample-preparation-and-confocal-laser-scanning-microscopy/ Fri, 02 May 2014 15:04:00 +0000 https://www.leica-microsystems.com/11000 Confocal Microscopy Neuroscience Live-Cell Imaging In mice, the ability to detect pheromones is principally mediated by the vomeronasal organ (VNO). Here, an acute tissue slice preparation of VNO for performing calcium imaging is described. This physiological approach allows observations of subpopulations and/or individual neurons in a living tissue and is convenient for receptor-ligand identification. https://www.leica-microsystems.com//science-lab/imaging-pheromone-sensing-in-a-mouse-vomeronasal-acute-tissue-slice-preparation/ Fri, 02 May 2014 14:24:00 +0000 https://www.leica-microsystems.com/13105 Surface Metrology Digital Microscopy Surface free energy (SFE) is a property which depends on the chemical state and roughness of the surface and it is necessary to develop a reliable method to evaluate SFE value on a small area, taking into account these two different contributions. Today contact angle methods are the most used and they allow to evaluate the global mean value of SFE on areas of mm2 size. https://www.leica-microsystems.com//science-lab/an-innovative-non-contact-method-to-determine-surface-free-energy-on-micro-areas/ Mon, 28 Apr 2014 16:41:00 +0000 https://www.leica-microsystems.com/13379 Super-Resolution Focus on Microscopy 2014 was held at the University of Sydney, Australia from April 13–16. Over 600 delegates and exhibitors attend this year’s FOM meeting. The program with attached one-page abstracts of these presentations is now available as PDF on the FOM homepage. https://www.leica-microsystems.com//science-lab/focus-on-microscopy-2014-abstracts-of-the-presentations-online-available/ Fri, 25 Apr 2014 11:58:00 +0000 Prof. Filip Braet, Prof. Guy Cox, Prof. G.J. (Fred) Brakenhoff https://www.leica-microsystems.com/12785 Surface Metrology Digital Microscopy Quality Assurance Acidic beverages, such as soft drinks (orange juice, and cola), or ethanol (whisky), can produce erosion of resin composites. The surface degradation of resin materials is related to the content and distribution of the fillers, the composition of the matrix resin, and the effect of silane surface treatment on the fillers. https://www.leica-microsystems.com//science-lab/surface-roughness-of-flowable-resin-composites-eroded-by-acidic-and-alcoholic-drinks/ Thu, 24 Apr 2014 20:51:00 +0000 https://www.leica-microsystems.com/13283 Super-Resolution https://www.leica-microsystems.com//science-lab/galleries/event-galleries/image-gallery-focus-on-microscopy-sydney-2014/ Tue, 15 Apr 2014 22:35:00 +0000 Scott Strothers, Claudia Müller https://www.leica-microsystems.com/13257 Multiphoton Microscopy Live-Cell Imaging Multi-channel multiphoton microscopy with dedicated optics for CLARITY. The glomerular filtration barrier (GFB) is a complex spatial structure within the kidney glomerulis where ultrafiltration takes place. Podocytes are critical elements of the GFB and take part in the filtration process. They have been shown to be involved in the development of kidney diseases. However, due to technical limitations, the mechanism of glomerular pathology is not well understood. https://www.leica-microsystems.com//science-lab/tracking-glomerular-fate-over-long-time-distances/ Mon, 14 Apr 2014 12:32:00 +0000 M.D., Ph.D. Janos Peti-Peterdi, PhD Isabelle Köster https://www.leica-microsystems.com/12790 Confocal Microscopy Live-Cell Imaging Fully automated tracking of moving 3D structures in living cells represents several challenges that are typical to the analysis of biological image data. Taking 4D image data from chloroplasts in living Elodea leaf cells as an example we demonstrate new approaches to meet these challenges. Chloroplasts are surprisingly dynamic organelles: They move extensively throughout cells, they regularly split or fuse and their shape changes constantly. https://www.leica-microsystems.com//science-lab/fully-automated-tracking-of-chloroplasts-in-elodea-leaf-cells-from-4d-image-data/ Wed, 09 Apr 2014 16:22:00 +0000 Dr. Johannes Zimmermann, Dr. Florian Leiss, Frank Sieckmann https://www.leica-microsystems.com/12779 Super-Resolution The widefield super-resolution technique GSDIM (Ground State Depletion followed by individual molecule return) is a localization microscopy technique that is capable of resolving details as small as 20 nanometers. GSDIM is suitable for a wide range of samples. https://www.leica-microsystems.com//science-lab/sample-preparation-for-gsdim-localization-microscopy-protocols-and-tips/ Tue, 01 Apr 2014 12:40:00 +0000 Dr. Marko Lampe, Dr. Christoph Greb https://www.leica-microsystems.com/13128 Super-Resolution Here we illustrate the protocol for imaging by two-color STED nanoscopy the cytotoxic immune synapse of NK cells recapitulated on glass. Using this method we obtain sub-100 nm resolution of synapse proteins and the cytoskeleton. https://www.leica-microsystems.com//science-lab/visualization-of-the-immunological-synapse-by-dual-color-time-gated-stimulated-emission-depletion-sted-nanoscopy/ Fri, 28 Mar 2014 16:34:00 +0000 Ph.D. Emily M. Mace, Ph.D., MD Jordan S. Orange https://www.leica-microsystems.com/2469 Laser Microdissection Neuroscience A characteristic sign of M. Parkinson is the deterioration of dopaminergic neurons in the mid-brain, specifically in the substantia nigra (SN, black substance). Different causes and forms of this disease have been identified. In the case of the genetic familial form, for example, it has been possible to identify various genes that have a causal influence for M. Parkinson. https://www.leica-microsystems.com//science-lab/the-morbus-parkinson-puzzle/ Thu, 27 Mar 2014 11:11:00 +0000 Dr. Falk Schlaudraff, Dr. Olaf Spörkel https://www.leica-microsystems.com/13103 Super-Resolution Stimulated emission depletion (STED) microscopy is a prominent approach of super-resolution optical microscopy, which allows cellular imaging with so far unprecedented unlimited spatial resolution. The introduction of time-gated detection in STED microscopy significantly reduces the (instantaneous) intensity required to obtain sub-diffraction spatial resolution. https://www.leica-microsystems.com//science-lab/a-new-filtering-technique-for-removing-anti-stokes-emission-background-in-gated-cw-sted-microscopy/ Tue, 25 Mar 2014 20:34:00 +0000 https://www.leica-microsystems.com/11782 EM Sample Preparation Chemical fixation of biological specimens for ultrastructural investigation is a relatively slow and selective process, and therefore a common source of artifacts. Freezing, on the other hand, is an excellent method to physically fix biological specimens in their entirety and without delay; the formation of ice crystals large enough to displace cellular material and destroy structures, would be, however, a major issue. https://www.leica-microsystems.com//science-lab/freeze-substitution-of-trypanosoma-brucei/ Tue, 18 Mar 2014 17:25:00 +0000 Harald Kotisch, Dr. Katy Schmidt, Dr. Jan Leunissen, Dr. Guenter Resch https://www.leica-microsystems.com/11150 Conservation The find of an almost life-sized sandstone statue of a Celtic "prince" in Glauberg in the German state of Hessen marked the beginning of an exciting journey to the early Celtic past of the 5th century BC, an age rich in myths and mystery due to the absence of written records. With their meticulous restoration of the valuable finds under the stereo microscope, restorers are helping to piece together the puzzle of our Celtic ancestors. https://www.leica-microsystems.com//science-lab/myths-under-the-microscope-the-restoration-of-celtic-finds/ Mon, 17 Mar 2014 17:27:00 +0000 https://www.leica-microsystems.com/12989 Confocal Microscopy DNA methylation plays a crucial role in development through inheritable gene silencing. Plants possess three types of DNA methyltransferases (MTases), namely Methyltransferase (MET), Chromomethylase (CMT) and Domains Rearranged Methyltransferase (DRM), which maintain methylation at CG, CHG and CHH sites. DNA MTases have not been studied in legumes so far. Here, we report the identification and analysis of putative DNA MTases in five legumes, including chickpea, soybean, pigeonpea, Medicago and Lotus. MTases in legumes could be classified in known MET, CMT, DRM and DNA nucleotide methyltransferases (DNMT2) subfamilies based on their domain organization. https://www.leica-microsystems.com//science-lab/genomic-survey-gene-expression-analysis-and-structural-modeling-suggest-diverse-roles-of-dna-methyltransferases-in-legumes/ Fri, 14 Mar 2014 11:29:00 +0000 https://www.leica-microsystems.com/12768 Neuroscience Confocal Microscopy Multiphoton Microscopy Imaging whole brains with CLARITY and multiphoton microscopy. Image a whole brain without sectioning? Investigate neuronal circuits without reconstruction? Perform molecular phenotyping without destroying subcellular structures? Understanding the brain with molecular resolution and global scope has always been challenging. The novel CLARITY method, developed by the Deisseroth laboratory at Stanford University, USA, pushes the barrier of deep tissue imaging a big step ahead. https://www.leica-microsystems.com//science-lab/map-the-brain-with-clarity/ Tue, 11 Mar 2014 07:26:00 +0000 PhD Isabelle Köster https://www.leica-microsystems.com/12418 Confocal Microscopy Live-Cell Imaging In endocrine cells within islets of Langerhans calcium ions couple cell stimulation to hormone secretion. Since the advent of modern fluorimetry, numerous in vitro studies employing primarily isolated mouse islets have investigated the effects of various secretagogues on cytoplasmic calcium, predominantly in insulin-secreting beta cells. Due to technical limitations, insights of these studies are inherently limited to a rather small subpopulation of outermost cells. https://www.leica-microsystems.com//science-lab/glucose-stimulated-calcium-dynamics-in-islets-of-langerhans-in-acute-mouse-pancreas-tissue-slices/ Tue, 11 Mar 2014 07:00:00 +0000 https://www.leica-microsystems.com/12420 Confocal Microscopy Live-Cell Imaging We propose a network representation of electrically coupled beta cells in islets of Langerhans. Beta cells are functionally connected on the basis of correlations between calcium dynamics of individual cells, obtained by means of confocal laser-scanning calcium imaging in islets from acute mouse pancreas tissue slices. https://www.leica-microsystems.com//science-lab/functional-connectivity-in-islets-of-langerhans-from-mouse-pancreas-tissue-slices/ Mon, 10 Mar 2014 10:31:00 +0000 https://www.leica-microsystems.com/12787 Laser Microdissection Many animal experiments have been performed to date to elucidate the mechanism of cardiac ischemic injury, and these studies have enhanced our understanding of cardiac remodeling following hypoxic stress. Clinical studies using samples from admitted patients have also been conducted to discover diagnostic or prognostic biomarkers for acute myocardial infarction (AMI). https://www.leica-microsystems.com//science-lab/sorbin-and-sh3-domain-containing-protein-2-is-released-from-infarcted-heart-in-the-very-early-phase-proteomic-analysis-of-cardiac-tissues-from-patients/ Wed, 05 Mar 2014 19:11:00 +0000 https://www.leica-microsystems.com/12772 Neuroscience Optogenetics is a technique that allows scientists to control neurons’ electrical activity with light by engineering them to express light-sensitive proteins. Within the past decade, it has become a very powerful tool for discovering the functions of different types of cells in the brain. https://www.leica-microsystems.com//science-lab/optogenetic-toolkit-goes-multicolor/ Tue, 25 Feb 2014 12:35:00 +0000 Anne Trafton https://www.leica-microsystems.com/12635 Quality Assurance In the automotive industry, the technical cleanliness of function-critical individual and system components has become an increasingly critical criterion for reliability and service life. This trend is also reflected in ISO/DIS 16232 (road vehicles – cleanliness of components of fluid circuits). Microscope systems with corresponding analytical software enable efficient and reliable residual dirt analysis of injectors, pumps, control units and other micromechanical components. https://www.leica-microsystems.com//science-lab/clean-parts-more-reliable-and-longer-lifetime/ Mon, 24 Feb 2014 12:56:00 +0000 Dr. Kay Scheffler, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/12648 Surface Metrology Quality Assurance Digital Microscopy Titanium (Ti) alloys are biocompatible materials which are often used for human implants, especially in dentistry. For this study, different types of odontological (dental) treatments for peri-implantitis, an infection of the gum and dental tissue around the implant, were performed on a Ti-6Al-4V (TAV) alloy, typically used for implants, to study the effect on its surface roughness and chemistry. https://www.leica-microsystems.com//science-lab/the-effects-of-peri-implantitis-decontamination-treatments-on-the-surface-roughness-and-chemistry-of-a-titanium-alloy-used-for-dental-implants-implications-for-bone-reintegration/ Wed, 19 Feb 2014 11:22:00 +0000 PhD Jordi Diaz-Marcos, MSc Joan Vilana Balastegui, DDS, MS, PhD Angel F. Espías Gómez, Dr. Luis Alberto Sánchez Soler, Dr. Frédéric Parahy https://www.leica-microsystems.com/12452 Neuroscience Live-Cell Imaging From neuroscience, biotechnology and digital media to sustainable energy and cloud computing, almost everything today is somehow affected – and sometimes entirely reshaped – by scientific and technological advances. https://www.leica-microsystems.com//science-lab/good-to-know/the-50-most-influential-scientists-in-the-world-today/ Mon, 17 Feb 2014 19:03:00 +0000 https://www.leica-microsystems.com/12426 Confocal Microscopy Neuroscience The cellular organization of the cortex is of fundamental importance for elucidating the structural principles that underlie its functions. It has been suggested that reconstructing the structure and synaptic wiring of the elementary functional building block of mammalian cortices, the cortical column, might suffice to reverse engineer and simulate the functions of entire cortices. https://www.leica-microsystems.com//science-lab/cellular-organization-of-cortical-barrel-columns-is-whisker-specific/ Fri, 14 Feb 2014 13:01:00 +0000 https://www.leica-microsystems.com/11035 Stereo Microscopy In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors while the second involves a wholesale (global) activation of most or all such neurons. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons. https://www.leica-microsystems.com//science-lab/local-and-global-methods-of-assessing-thermal-nociception-in-drosophila-larvae/ Fri, 14 Feb 2014 12:46:00 +0000 https://www.leica-microsystems.com/11772 EM Sample Preparation Seeds accumulate proteins and starch which will be broken down and mobilized upon germination. Among seeds, cereals constitute an example of a highly specialized storage tissue that constitutes up to 80% of the total seed volume (Watson 1987). The seed endomembrane system is highly specialized and seed storage proteins travel through the endomembrane system en route to the protein bodies, which are either derived from the Endoplasmic reticulum (ER) or of vacuolar origin (Muntz 1998). https://www.leica-microsystems.com//science-lab/protein-trafficking-in-cereal-seeds/ Mon, 10 Feb 2014 17:31:00 +0000 PhD Elsa Arcalis, PhD, MSc Eva Stöger https://www.leica-microsystems.com/12430 Confocal Microscopy With over a million described species arthropods, which include spiders, millipedes, crustaceans and insects, are the most diverse taxon on earth. The total number of species of these segmented armored animals is still unknown and many of them are still undiscovered. Scientists estimate the number of arthropods of a total of ten to thirty million species. https://www.leica-microsystems.com//science-lab/evolutionary-morphology-rostock-scientists-clarify-phylogenetic-relationships-within-the-most-diverse-taxon-arthropoda/ Thu, 06 Feb 2014 10:40:00 +0000 M. A. Janika Wiesner, Dr. Martin Fritsch https://www.leica-microsystems.com/11809 EM Sample Preparation Freeze-substitution is a process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient-temperature dehydration. During freeze substitution the “frozen” water is dissolved by an organic solvent, which usually also contains chemical fixatives. https://www.leica-microsystems.com//science-lab/brief-introduction-to-freeze-substitution/ Tue, 04 Feb 2014 09:32:00 +0000 Riet de Rycke, Dr. Andres Kaech https://www.leica-microsystems.com/12449 Laser Microdissection Neuroscience High-quality total RNA has been prepared from cell bodies of mouse spinal cord motor neurons by laser capture microdissection after staining spinal cord sections with Azure B in 70% ethanol. Sufficient RNA (~40–60 ng) is recovered from 3,000–4,000 motor neurons to allow downstream RNA analysis by RNA-seq and qRT-PCR. https://www.leica-microsystems.com//science-lab/production-of-rna-for-transcriptomic-analysis-from-mouse-spinal-cord-motor-neuron-cell-bodies-by-laser-capture-microdissection/ Thu, 30 Jan 2014 09:11:00 +0000 https://www.leica-microsystems.com/12284 Super-Resolution Super-resolution microscopy such as Stimulated Emission Depletion (STED) and single-molecule based techniques rely on the same principle for breaking the diffraction limit: the unwanted fluorescence signals are switched off during the image acquisition process. Consequently, Ground State Depletion followed by Individual Molecule Return (GSDIM) microscopy and related techniques like PALM, STORM and dSTORM use metastable dark states of a fluorophore for temporal separation of single molecules. https://www.leica-microsystems.com//science-lab/the-force-of-the-dark-side-embedding-media-for-gsdim-super-resolution-localization-microscopy/ Thu, 23 Jan 2014 18:49:00 +0000 Sebastian Bänfer https://www.leica-microsystems.com/12287 Fluorescence Microscopy Quantitative Imaging Live-Cell Imaging Small brightly fluorescent carbon nanoparticles have emerged as a new class of materials important for sensing and imaging applications. We analyze comparatively the properties of nanodiamonds, graphene and graphene oxide ‘dots’, of modified carbon nanotubes and of diverse carbon nanoparticles known as ‘C-dots’ obtained by different methods. https://www.leica-microsystems.com//science-lab/novel-fluorescent-carbonic-nanomaterials-for-sensing-and-imaging/ Thu, 16 Jan 2014 11:53:00 +0000 https://www.leica-microsystems.com/11169 Education "The lack of pathological examinations is one of the most pressing issues of medical care in developing countries." Dr. Roberto Monaco, pathologist in Neaples, knows the disastrous consequences from his own experience. He is a member of the board of the Italian NGO "Patologi oltre Frontiera" ("Pathologists beyond the Borders") and campaigns in this capacity for the implementation of pathological examinations as the basis for diagnosis and therapy even in the poorest countries. https://www.leica-microsystems.com//science-lab/pathology-overcomes-boundaries-interactive-microscope-system-facilitates-training-of-pathologists-in-developing-countries/ Fri, 10 Jan 2014 09:14:00 +0000 Dr. Roberto Monaco https://www.leica-microsystems.com/12002 Confocal Microscopy Live-Cell Imaging Oscillatory electrical activity is regarded as a hallmark of the pancreatic beta cell glucose-dependent excitability pattern. Electrophysiologically recorded membrane potential oscillations in beta cells are associated with in-phase oscillatory cytosolic calcium activity ([Ca2+]i) measured with fluorescent probes. Recent high spatial and temporal resolution confocal imaging revealed that glucose stimulation of beta cells in intact islets within acute tissue slices produces a [Ca2+]i change with initial transient phase followed by a plateau phase with highly synchronized [Ca2+]i oscillations. https://www.leica-microsystems.com//science-lab/the-relationship-between-membrane-potential-and-calcium-dynamics-in-glucose-stimulated-beta-cell-syncytium-in-acute-mouse-pancreas-tissue-slices/ Tue, 17 Dec 2013 17:08:00 +0000 https://www.leica-microsystems.com/11802 Super-Resolution The Nuclear Pore Complex (NPC) is a large complex in the nuclear membrane, representing the gate to the eukaryotic genetic makeup. Because of this outstanding function the structure of the NPC is of great interest. Anna Szymborska, scientist at the EMBL in Heidelberg, comments on her resaerch results and the potential of Ground State Depletion microscopy (GSD) for protein complex analysis in the following interview. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-gives-new-insights-into-nuclear-pore-complex-organization/ Wed, 11 Dec 2013 17:53:00 +0000 Dr. Anna Szymborska, Dr. Jan Ellenberg, Dr. Christoph Greb https://www.leica-microsystems.com/11133 Laser Microdissection New and far-reaching applications have recently been opened up in the field of laser microdissection. In addition to conventional dissection, the laser microdissection system (LMD) of Leica Microsystems is an excellent tool for marking relevant structures, offering highly specific laser manipulation of selected areas. This laser marking function is useful for applications such as CLEM, NanoSIMS as well as in the live cell sector. https://www.leica-microsystems.com//science-lab/laser-microdissection-lmd-and-fancy-applications/ Mon, 09 Dec 2013 13:02:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/11693 Live-Cell Imaging Super-Resolution Confocal Microscopy Neuroscience Fluorescence Microscopy Since its introduction in the 1600s, improvements in microscope technology have continually broadened the types of cells and cellular processes that can be studied. Advances in automation have made this already-simple tool faster and more capable, and time-lapse imaging reveals function and dynamics in addition to structure. Live-cell imaging has enabled us to witness incredible moments in biology in unprecedented detail. Even embryogenesis – the process of cell division and cellular differentiation that occurs at the earliest stages of life – has recently been captured. https://www.leica-microsystems.com//science-lab/live-cell-imaging-evolves-to-find-new-niches/ Mon, 25 Nov 2013 10:15:00 +0000 https://www.leica-microsystems.com/10668 Laser Microdissection This video explains the laser beam deflection principle of Leica Microsystems‘ laser microdissection systems (Leica LMD). The laser, the beam expander, motorized attenuator, motorized iris diaphragm for limiting the aperture for cutting and the scan unit are situated in the instrument’s special incident light axis. The scan unit consists of two prism wedges whose thickness and wedge angle are chosen so that the laser beam, after passing through the two prism wedges, is directed precisely through the center of the back focal plane of the objective. https://www.leica-microsystems.com//science-lab/video-how-to-achieve-fast-precise-cutting-lines-in-laser-microdissection/ Mon, 18 Nov 2013 14:32:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/11137 Live-Cell Imaging Confocal Microscopy Neuroscience Euro‐BioImaging is a large‐scale pan‐European research infrastructure project on the European Strategy Forum on Research Infrastructures (ESFRI) Roadmap. Its mission is to build a distributed imaging infrastructure across Europe that will provide open access to innovative biological, molecular and medical imaging technologies for European researchers. https://www.leica-microsystems.com//science-lab/good-to-know/euro-bioimaging-the-pan-european-open-access-research-infrastructure-project-for-biological-and-medical-imaging-technologies/ Wed, 13 Nov 2013 11:03:00 +0000 M.Sc., Cornelia Gilbrich-Wille, Dr. Tanja Ninkovic https://www.leica-microsystems.com/11119 Stereo Microscopy With over 35,000 known species, ground beetles – or Carabidae – are among the most speciose groups of animals in the world. Biologist Dr. Joachim Schmidt devotes his entire scientific work to the research of these frequently very small beetles, their ecology, distribution and phylogeny. He is particularly interested in the ground beetles of high mountain regions. https://www.leica-microsystems.com//science-lab/ground-beetles-shed-light-on-the-environmental-history-of-high-mountain-regions/ Tue, 05 Nov 2013 12:48:00 +0000 M. A. Janika Wiesner, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/11089 Super-Resolution Optical imaging devices have a finite depth of field and diffraction limited resolution. The depth of field problem was tackled first with confocal microscopes, diffraction unlimited resolution is available since a few years with super-resolution microscopes. Super-resolution microscopes with a solved depth of field problem are now available. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-and-the-third-dimension/ Mon, 04 Nov 2013 09:55:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/11235 Confocal Microscopy Acousto-optical elements have successfully replaced planar filters in many positions. The white confocal, regarded as the fully spectrally tunable confocal microscope, was not possible without this technique. Acousto-optical elements are highly transparent, quickly tunable and allow many colors to be managed simultaneously. As they show a strong dependence in polarization and have comparably small dimensions, their active part is used to modify and guide the laser illumination light, thereby leaving the principal beam (0th order) unaffected. Excitation color selection and attenuation (excitation filtering), as well as separation of illumination and detection light (beam splitting) are the main fields of application. https://www.leica-microsystems.com//science-lab/acousto-optics-in-true-confocal-spectral-microscope-systems/ Fri, 01 Nov 2013 05:47:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/10717 EM Sample Preparation Immunoelectron microscopy is a powerful method to diagnose viral diseases and to study the distribution of the viral agent within plant cells and tissues. Nevertheless, current protocols for the immunological detection of viral diseases with transmission electron microscopy (TEM) in plants take between 3 and 6 days and are therefore not suited for rapid diagnosis of virus diseases in plants. In this study, we describe a method that allows rapid cytohistochemical detection of tobacco mosaic virus (TMV) in leaves of tobacco plants. https://www.leica-microsystems.com//science-lab/rapid-immunohistochemical-diagnosis-of-tobacco-mosaic-virus-disease-by-microwave-assisted-plant-sample-preparation/ Wed, 16 Oct 2013 20:36:00 +0000 Prof. Günther Zellnig, Stefan Möstl, Prof. Bernd Zechmann https://www.leica-microsystems.com/11033 Neuroscience Stereo Microscopy Live-Cell Imaging Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact. https://www.leica-microsystems.com//science-lab/patch-clamp-recordings-from-embryonic-zebrafish-mauthner-cells/ Fri, 11 Oct 2013 21:40:00 +0000 https://www.leica-microsystems.com/11038 Confocal Microscopy Live-Cell Imaging On October 7th 2013, The Nobel Assembly at Karolinska Institutet has decided to award The Nobel Prize in Physiology or Medicine 2012 jointly to James E. Rothman, Randy W. Schekman and Thomas C. Südhof "for their discoveries of machinery regulating vesicle traffic, a major transport system in our cells". https://www.leica-microsystems.com//science-lab/nobel-prize-2013-in-physiology-or-medicine-for-discoveries-of-the-machinery-regulating-vesicle-traffic/ Fri, 11 Oct 2013 15:14:00 +0000 https://www.leica-microsystems.com/10734 EM Sample Preparation Employing liquid nitrogen (LN2) as a coolant in the complex process of high pressure freezing raises certain considerations regarding phase transition not only of the liquid sample to be frozen but also of the element used to freeze the sample. According to the phase diagram of nitrogen, changes in temperature or pressure will alter the state of this element. At ambient pressure nitrogen is liquid at temperatures between –209,86 °C and –195,80 °C. It is a rather fragile equilibrium within limited phase boundaries. https://www.leica-microsystems.com//science-lab/thermodynamic-considerations-regarding-the-ln2-in-a-high-pressure-freezer/ Thu, 10 Oct 2013 21:05:00 +0000 Dr. Paul Wurzinger, Dr. Cveta Tomova https://www.leica-microsystems.com/11024 Super-Resolution Stefan Hell is credited with having conceived and applied the first viable concept for breaking Abbe’s diffraction limit in a light-focusing microscope. He is a scientific member of the Max Planck Society and a director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, where he currently leads the department of nanobiophotonics. https://www.leica-microsystems.com//science-lab/interview-with-stefan-hell-breaking-nanoscale-boundaries/ Wed, 09 Oct 2013 13:29:00 +0000 Dr. Peter Andersen https://www.leica-microsystems.com/10924 Confocal Microscopy Live-Cell Imaging Constructing a body is like building a house – if you compromise structural integrity, the edifice can collapse. Nowhere is that clearer on a cellular level than in the case of epithelial sheets, single layers of cells that line every body cavity from the gut to mammary glands. Stowers Institute for Medical Research Associate Investigator Matt Gibson, PhD, and his team use simple animal systems like fruit flies and sea anemones to investigate how epithelial cells maintain order while getting jostled by cell division. https://www.leica-microsystems.com//science-lab/a-flip-of-the-mitotic-spindle-has-disastrous-consequences-for-epithelial-cells/ Fri, 04 Oct 2013 07:58:00 +0000 PhD Yu-ichiro Nakajima, PhD Matthew C. Gibson https://www.leica-microsystems.com/10912 EM Sample Preparation Since the contrast in the electron microscope depends primarily on the differences in the electron density of the organic molecules in the cell, the efficiency of a stain is determined by the atomic weight of the stain attached to the biological structures. Consequently, the most widely used stains in electron microscopy are the heavy metals uranium and lead. https://www.leica-microsystems.com//science-lab/brief-introduction-to-contrasting-for-em-sample-preparation/ Wed, 02 Oct 2013 14:33:00 +0000 Dr. Ruwin Pandithage https://www.leica-microsystems.com/10864 Ergonomics Stereo Microscopy Quality Assurance Sensor and identification technology, industrial cameras: when top precision is required, the name Baumer Electric is soon mentioned. The Swiss company designs and produces components for manufacturing facilities all over the world. To be able to guarantee that the finished components are of the highest possible quality, elaborate quality inspections are carried out part for part under the stereo microscope. https://www.leica-microsystems.com//science-lab/higher-motivation-longer-concentration-ergonomics-as-a-competitive-advantage/ Tue, 01 Oct 2013 09:38:00 +0000 Michael Birlenbach, Remo Holenstein https://www.leica-microsystems.com/10826 Super-Resolution When Stefan Hell invented a technique that pushed beyond the diffraction limit of light for the first time, a new field of nanoscale imaging was born. Now laboratories around the world are using STED microscopy to smash through more boundaries in fundamental science. https://www.leica-microsystems.com//science-lab/sted-microscopy-a-new-chapter-in-light-imaging/ Mon, 16 Sep 2013 10:28:00 +0000 https://www.leica-microsystems.com/10224 EM Sample Preparation Glass knives are used in an ultramicrotome to cut ultrathin slices of samples for electron and light microscope applications. For resin and for cryo sections (Tokuyasu samples) the knife edge must be extremely sharp, strong and stable. An important requirement for breaking glass knives of superior quality is the quality of the glass strips. Glass strips are produced from specially selected glass, the thickness and quality of which is precisely controlled. Only strict tolerances ensured by careful quality control allow breaking of two high quality knives from one square. https://www.leica-microsystems.com//science-lab/brief-introduction-to-glass-knifemaking-for-electron-and-light-microscope-applications/ Fri, 13 Sep 2013 15:49:00 +0000 Dr. Ruwin Pandithage https://www.leica-microsystems.com/10792 Ergonomics To do their best at work, people have to have a feeling of physical wellbeing: they are more attentive and motivated and more productive for longer periods of time. As ergonomically designed workplaces are conducive to physical wellbeing, ergonomics makes a direct contribution to the operating result, reducing sick days and enhancing productivity. https://www.leica-microsystems.com//science-lab/ergonomics/investing-in-ergonomically-designed-microscope-workplaces-pays-off/ Fri, 06 Sep 2013 07:18:00 +0000 Claudia Müller, John Ludescher https://www.leica-microsystems.com/10555 Super-Resolution With the new 3D GSDIM technique structures like the Golgi and the microtubular network are resolved not only laterally, but also in a third dimension. The principle is based on the use of optical astigmatism to determine the accurate lateral and axial position of individual fluorochromes. https://www.leica-microsystems.com//science-lab/three-dimensional-super-resolution-gsdim-microscopy/ Wed, 04 Sep 2013 15:24:00 +0000 PhD Tamara Straube, Sebastian Bänfer, Prof. Ralf Jacob https://www.leica-microsystems.com/10730 Super-Resolution Confocal Microscopy Although deconvolution can improve the quality of any type of microscope, the high computational time required has so far limited its massive spreading. Here we demonstrate the ability of the scaled-gradient-projection (SGP) method to provide accelerated versions of the most used algorithms in microscopy. To achieve further increases in efficiency, we also consider implementations on graphic processing units (GPUs). https://www.leica-microsystems.com//science-lab/towards-real-time-image-deconvolution-application-to-confocal-and-sted-microscopy/ Mon, 02 Sep 2013 07:51:00 +0000 https://www.leica-microsystems.com/10222 EM Sample Preparation Coating of samples is required in the field of electron microscopy to enable or improve the imaging of samples. Creating a conductive layer of metal on the sample inhibits charging, reduces thermal damage and improves the secondary electron signal required for topographic examination in the SEM. https://www.leica-microsystems.com//science-lab/brief-introduction-to-coating-technology-for-electron-microscopy/ Wed, 28 Aug 2013 12:10:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/10447 Laser Microdissection Live-Cell Imaging EM Sample Preparation Confocal Microscopy To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins with electron microscopy. With laser micro-patterned culture substrate, we created coordinates that were conserved at every step of the sample preparation and visualization processes. Specifically designed for cryo-fixation, this method allowed a fast freezing of dynamic events within seconds and their ultrastructural characterization. https://www.leica-microsystems.com//science-lab/from-dynamic-live-cell-imaging-to-3d-ultrastructure-novel-integrated-methods-for-high-pressure-freezing-and-correlative-light-electron-microscopy/ Wed, 28 Aug 2013 11:41:00 +0000 https://www.leica-microsystems.com/10216 Super-Resolution Multiphoton Microscopy Two-photon microscopy, multiphoton microcopy and super-resolution imaging. We developed a new class of two-photon excitation–stimulated emission depletion (2PE-STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging, exciting and stimulating the emission of a fluorophore by means of a single wavelength. https://www.leica-microsystems.com//science-lab/single-wavelength-two-photon-excitation-stimulated-emission-depletion-sw2pe-sted-superresolution-imaging/ Fri, 23 Aug 2013 11:02:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/10653 Super-Resolution Live-Cell Imaging The newly developed STED-RICS microscopy method records rapid movements of molecules in live samples. By combining raster image correlation spectroscopy (RICS) with STED fluorescence microscopy, researchers of Karlsruhe Institute of Technology (KIT) opened up new applications in medical research, e.g. analyzing the dynamics of cell membranes at high protein concentrations. https://www.leica-microsystems.com//science-lab/watching-molecule-movements-in-live-cells/ Thu, 22 Aug 2013 13:28:00 +0000 https://www.leica-microsystems.com/9350 Fluorescence Microscopy Confocal Microscopy Live-Cell Imaging Fluorescence microscopy and other light-based applications require optical filters that have demanding spectral and physical characteristics. Often, these characteristics are application-specific and an optic that might be appropriate and optimal for one is both inappropriate and sub-optimal for another. https://www.leica-microsystems.com//science-lab/handbook-of-optical-filters-for-fluorescence-microscopy/ Tue, 06 Aug 2013 07:59:00 +0000 BSc. Jay Reichman https://www.leica-microsystems.com/10290 Super-Resolution Live-Cell Imaging Neuroscience Since super-resolution microscopy techniques revolutionized the concept of light microscopy by overcoming the physical diffraction limit, STED microscopy and other super-resolution techniques have aroused considerable interest. The diffraction limit imposes no more constraints on resolution. New microscopes with ever-decreasing resolution limits are being developed, for instance by the inventor of STED microscopy, Prof. Stefan Hell, now director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany. https://www.leica-microsystems.com//science-lab/new-labeling-tools-can-help-to-realize-the-full-potential-of-super-resolution-microscopy/ Fri, 02 Aug 2013 10:55:00 +0000 Dr. Matthias Schauen, Dr. Felipe Opazo, Prof. Silvio Rizzoli https://www.leica-microsystems.com/10062 EM Sample Preparation Especially in core EM-facilities one is confronted with material (microorganisms and cells) which are infectious. It is a must to follow the safety rules according to the National regulations. However even when safe laboratories are available it is very convenient to know that the applied high pressure freezing system is in itself safe. https://www.leica-microsystems.com//science-lab/safe-high-pressure-freezing-of-infectious-microorganisms/ Wed, 24 Jul 2013 14:10:00 +0000 PhD Dimitri Vanhecke, Dr. Benoit Zuber, Dr. Silvio D. Brugger, Daniel Studer https://www.leica-microsystems.com/10281 CLEM EM Sample Preparation Live-Cell Imaging Confocal Microscopy CLEM (correlative live cell and electronmicroscopy) seeks to bridge the data acquired with different imaging strategies, typically between light microscopy and electron microscopy. It has been successfully applied in cell cultures, although its use in multicellular systems is hampered by difficulties in locating the ROI (region of interest). https://www.leica-microsystems.com//science-lab/a-precise-and-rapid-mapping-protocol-for-correlative-light-and-electron-microscopy-of-small-invertebrate-organisms/ Tue, 16 Jul 2013 09:26:00 +0000 https://www.leica-microsystems.com/10286 Live-Cell Imaging Multiphoton Microscopy Cancer research using multiphoton microscopy. High-grade gliomas (World Health Organization grade III anaplastic astrocytoma and grade IV glioblastoma multiforme), the most prevalent primary malignant brain tumors, display a cellular hierarchy with self-renewing, tumorigenic cancer stem cells (CSCs) at the apex. While the CSC hypothesis has been an attractive model to describe many aspects of tumor behavior, it remains controversial due to unresolved issues including the use of ex vivo analyses with differential growth conditions. https://www.leica-microsystems.com//science-lab/direct-in-vivo-evidence-for-tumor-propagation-by-glioblastoma-cancer-stem-cells/ Fri, 12 Jul 2013 13:21:00 +0000 https://www.leica-microsystems.com/10068 EM Sample Preparation The coating layers applied and used for electron microscopy imaging are commonly controlled and measured by quartz crystals. These crystals oscillate with a certain frequency (around 6 megahertz when new). By measuring the frequency before coating and after coating, the specific weight of the coating material and the geometric position of the quartz, the applied thickness can be calculated. https://www.leica-microsystems.com//science-lab/carbon-thickness-evaluation-in-electron-microscopy/ Thu, 11 Jul 2013 09:10:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/10135 Super-Resolution Widefield Microscopy The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and widefield super-resolution users from ten European countries took three days’ out to deepen their knowledge on super-resolution techniques and applications and make use of an opportunity for full exchange of experiences. https://www.leica-microsystems.com//science-lab/abstracts-of-the-3rd-european-super-resolution-user-club-meeting/ Mon, 08 Jul 2013 10:27:00 +0000 Prof. Alberto Diaspro, Ph.D. Illaria Testa, Prof. Silvio Rizzoli, Ph.D. Giuseppe Vicidomini, Prof. Ralf Jacob, Ph.D. Eric Hosy, Prof. Colin Sheppard, Zeno Lavagnino https://www.leica-microsystems.com/10208 Super-Resolution In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-with-time-gated-detection-theoretical-and-experimental-aspects/ Mon, 08 Jul 2013 08:48:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/9449 Surgical Microscopy Ergonomics Operations not only demand maximum concentration of the surgeon but also maximum physical fitness. Physiotherapist John Ludescher explains how the body can be given optimal support to perform at its best. The Austrian qualified as physiotherapist in Switzerland and has had his own practice for nine years. He is the author of the book “Aufrecht – Bewusst – Stark” (Upright – Aware – Strong) and designer of the “PhysioCap”. https://www.leica-microsystems.com//science-lab/ergonomically-designed-surgical-microscopes-support-performance/ Mon, 01 Jul 2013 17:41:00 +0000 John Ludescher https://www.leica-microsystems.com/10147 Confocal Microscopy To specifically collect emission from multiple probes, the light is first separated spatially and then passes through a device that defines a spectral band. Classically, this is a common glass-based bandpass filter. More recent approaches employ arrays for fixed-band detection or moving mirror sliders for fully tunable band characteristics. https://www.leica-microsystems.com//science-lab/spectral-detection-how-to-define-the-spectral-bands-that-collect-probe-specific-emission/ Thu, 27 Jun 2013 18:21:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/10106 Confocal Microscopy Stereo Microscopy Fluorescence Microscopy Spectacular discoveries in cardiac tissue regeneration are rapidly moving researchers closer to the goal of harnessing regenerative techniques to repair the human heart. Only eleven years ago, Dr. Kenneth Poss, Professor of Cell Biology at Duke University and an Early Career Scientist of the Howard Hughes Medical Institute, published the first research to clearly visualize an example of cardiac tissue regeneration using fluorescence microscopy. https://www.leica-microsystems.com//science-lab/organ-regeneration-an-unlikely-fish-tale/ Thu, 20 Jun 2013 08:01:00 +0000 PhD Katharine H. Hendrix https://www.leica-microsystems.com/10111 Confocal Microscopy Lasers have been critical to the advancement on confocal microscopy, and the white light laser (WLL) offers particular advantages. Finessing WLL output for bioimaging is a complex task, though, and traditional approaches retain key limitations. But acousto-optical beamsplitting enables smoother operation, leading to enhanced microscopy capabilities. https://www.leica-microsystems.com//science-lab/multiple-microscopy-modes-in-a-single-sweep-with-supercontinuum-white-light/ Fri, 14 Jun 2013 14:09:00 +0000 Dr. Lioba Kuschel, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/10060 EM Sample Preparation CLEM Water is the most abundant cellular constituent and therefore important for preserving cellular ultra-structure. Currently the only way to fix cellular constituents without introducing significant structural alterations is by cryo-fixation. There are currently two common methods employed; plunge freezing and high pressure freezing. https://www.leica-microsystems.com//science-lab/brief-introduction-to-high-pressure-freezing/ Wed, 12 Jun 2013 17:59:00 +0000 Dr. Cveta Tomova https://www.leica-microsystems.com/9680 Confocal Microscopy Square and hexagonal pinholes provide identical image signal levels, if the geometries are compared in a sensible manner. The amount of light passing the pinhole depends on the area of that aperture, consequently the area is the parameter that must be compared when discussing brightness of focus images. The use of incommensurable edge lengths is meant to confuse the reader and thus dishonest and reprehensible. In this article, the signal level as a function of geometry and size in confocal microscopes is described. https://www.leica-microsystems.com//science-lab/pinhole-geometry-four-corners-are-perfect/ Thu, 23 May 2013 12:59:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/9759 Quantitative Imaging Fluorescence Recovery After Photobleaching (FRAP) has been considered the most widely applied method for observing translational diffusion processes of macromolecules. The resulting information can be used to determine kinetic properties, like the diffusion coefficient, mobile fraction, and transport rate of the fluorescently labeled molecules. FRAP employs irradiation of a fluorophore with a short laser pulse. State of the art laser scanning microscopes, like the TCS SP8 confocal system, have the advantage of using a high intensity laser for photobleaching and a low intensity laser for image recording. With the LAS AF application wizard you can choose between different ways to carry out a FRAP experiment. https://www.leica-microsystems.com//science-lab/step-by-step-guide-for-frap-experiments/ Fri, 17 May 2013 13:50:00 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/13385 Surface Metrology The precise measurement of surface structures and topography is the key thing during the production, control and development in many sectors of industry or research. Because very often there is not possible to use some contact methods for performing this task new optical methods based on interferometry and confocal technology started to be available for non-contact surface metrology in the recent years. https://www.leica-microsystems.com//science-lab/automated-digital-3d-topography-measurement-with-the-leica-dcm-3d-dual-core-measuring-microscope/ Fri, 17 May 2013 09:39:00 +0000 https://www.leica-microsystems.com/9585 Laser Microdissection Modern laser microdissection technology has its roots in the early 20th century. It has been steadily advanced and modified over the years. The following article is a summary of the history of laser microdissection from its origins to today’s state of the art. https://www.leica-microsystems.com//science-lab/history-of-laser-microdissection/ Mon, 06 May 2013 19:38:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/9571 Forensics It has always been a deeply rooted human need to give life a meaning after death. At the end of the 16th century, the friars of the Capuchin monastery in Palermo, Italy, began preserving corpses by embalming them. https://www.leica-microsystems.com//science-lab/in-the-catacombs-of-the-capuchin-monastery-in-palermo/ Thu, 02 May 2013 12:51:00 +0000 M. A. Janika Wiesner, Dipl. Biol. Kristina Baumjohann, Dr. Mark Benecke, M. A. Jörg Scheidt https://www.leica-microsystems.com/9604 Basics in Microscopy April 26th 2013 is the 170th anniversary of the birth of Ernst Leitz I (1843–1920), who turned a small optical workshop in Wetzlar into an optics company of world renown. At the beginning of the 20th century, the Ernst Leitz Optical Works rapidly became one of the leading suppliers of microscopes and optical products. An exhibition in Wetzlar’s New Town Hall looks back on his life and the milestones of his work. His business skills and his outstanding commitment to his employees and society in general made Ernst Leitz I a sovereign in the best sense of the word: He really was a person who “stood above it all” – not because he disregarded everyone else, but because we simply have to take our hats off to this exceptional personality whose work and actions are far greater than the general notion of a lifetime achievement. https://www.leica-microsystems.com//science-lab/the-entrepreneur-ernst-leitz-i/ Wed, 24 Apr 2013 00:47:00 +0000 Dr. Ralf Christofori https://www.leica-microsystems.com/9555 EM Sample Preparation Immersion freezing of thin aqueous specimens is an essential preparation technique for cryo-transmission electron microscopy (cryo-TEM), aiming to preserve fragile biological structures such as molecules and cells in their hydrated environment for a close-to-native visualization. For successful experiments, vitreous ice must be produced, surface contamination must be avoided, and, most important, the natural state of the structure must be preserved. https://www.leica-microsystems.com//science-lab/immersion-freezing-of-suspended-particles-and-cells-for-cryo-electron-microscopy/ Fri, 12 Apr 2013 11:48:00 +0000 Dr. Guenter Resch, Marlene Brandstetter, Lisa Königsmaier, Edith Urban, Angela M. Pickl-Herk https://www.leica-microsystems.com/9512 Basics in Microscopy April 26th 2013 is the 170th anniversary of the birth of Ernst Leitz I (1843-1920), who turned a small optical workshop in Wetzlar into an optics company of world renown. At the beginning of the 20th century, the Ernst Leitz Optical Works rapidly became one of the leading suppliers of microscopes and optical products. This history is illuminated by the two films presented here. https://www.leica-microsystems.com//science-lab/videos-on-the-leica-history-german-only/ Wed, 10 Apr 2013 20:21:00 +0000 https://www.leica-microsystems.com/9492 Live-Cell Imaging Fluorescence Microscopy Quantitative Imaging For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of these proteins in intact animals, tissues, and cells. For enzymes, imaging their activity also provides information on their function or functions, which does not necessarily correlate with their location. Metabolic mapping enables imaging of activity of enzymes. https://www.leica-microsystems.com//science-lab/imaging-enzymes-at-work/ Fri, 05 Apr 2013 14:27:00 +0000 https://www.leica-microsystems.com/9495 Basics in Microscopy Microscopy-based imaging is booming and the need for tools to retrieve quantitative data from images is urgent. This book provides simple but reliable tools to generate valid quantitative gene expression data, at the mRNA, protein and activity level, from microscopic images in relation to structures in cells, tissues and organs in 2D and 3D. https://www.leica-microsystems.com//science-lab/image-cytometry/ Thu, 04 Apr 2013 20:08:00 +0000 https://www.leica-microsystems.com/9410 Stereo Microscopy Fluorescence Microscopy Especially in fluorescence microscopy, excitation light is friend and foe in one. On the one hand, energy-rich excitation via a specific light wavelength of the fluorochrome resulting in a bright positive fluorochrome signal is highly welcome. On the other hand, "noise" caused by reflections of excitation light passing through the surfaces of optical elements needs to be extremely slight to generate a perfect black background. This relation is described as "signal-to-noise ratio", which is highly relevant for differentiating optically between fluorescence positive and negative cells. https://www.leica-microsystems.com//science-lab/stereo-microscopes-with-triplebeam-technology/ Tue, 02 Apr 2013 12:35:00 +0000 Björn Fuchs https://www.leica-microsystems.com/9374 Coherent Raman Scattering (CRS) Fluorescence microscopy assumed a pivotal role in cell biology once it was possible to stain cell components selectively by fluorescing dyes. One of the first explorers of targeted stainings, Paul Ehrlich, had the idea that something that stains specifically should also kill specifically – which was associated with the term “magic bullet”, the essential idea of chemotherapy. His group discovered Salvarsan, a tailored drug against syphilis – though not specific enough not to cause substantial side effects. Screening many fluorescent dyes led to a long list of stainings which are used in histology, including dyes like DAPI or hematoxylin and eosin. https://www.leica-microsystems.com//science-lab/unmask-the-hidden-without-probes-cars/ Thu, 21 Mar 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Stefanie Degenhartt https://www.leica-microsystems.com/9356 Live-Cell Imaging Stereo Microscopy Intravital imaging of inflammatory and remodeling processes has been an important area of research and has also motivated the creation of numerous transgenic reporter mouse models that express fluorescent proteins. Our paper (Kilarski, Güç et al. 2013) describes a new in vivo imaging technique, which design was based on innovative concept of using immunostaining for live cells and tissue structures on surgically exposed mouse dermis without causing harmful immunotoxic effect. https://www.leica-microsystems.com//science-lab/intravital-immunofluorescence-for-visualizing-the-microcirculatory-and-immune-microenvironments-in-the-mouse-ear-dermis/ Mon, 18 Mar 2013 23:00:00 +0000 PhD Witold W. Kilarski, Esra Güç, PhD Melody A. Swartz https://www.leica-microsystems.com/9287 EM Sample Preparation P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers. https://www.leica-microsystems.com//science-lab/targeting-of-peroxisomal-matrix-proteins-in-the-diatom-phaeodactylum-tricornutum/ Sun, 17 Mar 2013 23:00:00 +0000 Dr. Andreas Klingl https://www.leica-microsystems.com/8389 Forensics Stereo Microscopy There is no crime without clues. They may be obvious, like a cartridge case at the scene of the crime or clear signs of crowbar damage on a door. But sometimes, clues are microscopically small. Besides the classic fingerprints, perpetrators also leave hairs or fiber traces. https://www.leica-microsystems.com//science-lab/every-clue-counts-forensics-inconceivable-without-microscopy/ Wed, 06 Mar 2013 23:00:00 +0000 M. A. Janika Wiesner https://www.leica-microsystems.com/9043 Super-Resolution This article gives an overview of two different types of superresolution techniques. Stimulated emission depletion (STED) microscopy is a versatile and fast method that is based on point scanning microscopy – usually an extension of a confocal microscope. Ground state depletion imaging (GSDIM) is a parallel recording widefield approach that explores inherent switching of fluorochromes and typically comes with a TIRF microscope. The two methods use very different approaches to reach the same goal: to see more details in light microscopes than possible when diffraction limited. https://www.leica-microsystems.com//science-lab/sted-and-gsdim-diffraction-unlimited-resolution-for-all-types-of-fluorescence-imaging/ Thu, 28 Feb 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/8119 Confocal Microscopy This article summarizes the development and differences in design and functionality of confocal technology as far as spectral properties are concerned, from classical filter-based excitation and emission color selection to fully flexible spectral excitation and emission tuning. All three major components: light source with excitation color selection, beam splitting for incident illumination and detector emission filtering have been completely transformed. https://www.leica-microsystems.com//science-lab/the-white-confocal-spectral-gaps-closed/ Sun, 24 Feb 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Lioba Kuschel https://www.leica-microsystems.com/7981 Confocal Microscopy Modern biomedical research is currently dominated by imaging and measuring with optical microscopes. One branch of the microscopy technology is confocal microscopy. For correlation purposes, multiparameter fluorescence imaging is particularly of unique interest. This article is concerned with the spectral performance of the various modules in a confocal point-scanning microscope ("True Confocal System"), and how these modules have evolved to allow for tunability and flexibility in excitation and emission collection in multiple bands (channels). https://www.leica-microsystems.com//science-lab/the-white-confocal/ Tue, 19 Feb 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/8121 Confocal Microscopy Multiphoton Microscopy This article discusses detectors (more precisely: sensors), that are employed in single point, i.e. true confocal scanning microscopes. The sensors in such systems are usually photomultiplier tubes. Also, the silicon pendants of PMTs are used for particular applications, especially single-molecule measurements. A new development has led to chimeric devices called hybrid detector (HyD) which unite benefits of both technologies. https://www.leica-microsystems.com//science-lab/detectors-for-sensitive-detection-hyd/ Tue, 19 Feb 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus, Holger Birk, Frank Schreiber https://www.leica-microsystems.com/7401 EM Sample Preparation Neurotransmitter receptors and ion channels in the central nervous system are localized to synaptic and extrasynaptic membrane compartments of pre- and postsynaptic elements of neurons. The impact of the activation of these proteins on synaptic integration and regulation of transmitter release depends on their precise location relative to synapses, as well as on the density and coupling of molecules in microcompartments of the cells. High-resolution qualitative and quantitative visualization of membranebound receptors and ion channels is, therefore, essential for understanding their roles in cell communication. https://www.leica-microsystems.com//science-lab/capturing-neurotransmitter-receptors-and-ion-channels/ Thu, 14 Feb 2013 23:00:00 +0000 Daniel Althof, Akos Kulik https://www.leica-microsystems.com/8109 Confocal Microscopy Stereo Microscopy Live-Cell Imaging The human nervous system is an infinitely complex network consisting of some 100 billions of neurons. It is the result of many-faceted evolutionary processes spanning millions of years which, like the development of other organ systems, have been little researched so far. https://www.leica-microsystems.com//science-lab/molecular-developmental-biology-norwegian-marine-research-scientists-solve-the-mysteries-of-evolution/ Mon, 11 Feb 2013 23:00:00 +0000 M. A. Janika Wiesner, Dr. Andreas Hejnol https://www.leica-microsystems.com/7405 EM Sample Preparation Contrast in transmission electron microscopy (TEM) is mainly produced by electron scattering at the specimen: Structures that strongly scatter electrons are referred to as electron dense and appear as dark areas in the bright fi eld image, while structures which scatter fewer electrons appear bright (electron transparent). https://www.leica-microsystems.com//science-lab/perusing-alternatives-for-automated-staining-of-tem-thin-sections/ Mon, 04 Feb 2013 23:00:00 +0000 Nicole Fellner, Marlene Brandstetter, Karin Trimmel, Dr. Guenter Resch https://www.leica-microsystems.com/5726 EM Sample Preparation Cathodoluminescence microanalysis is an emerging technique that is fast gaining popularity in the world of materials science. CL is a light emission phenomena resulting from the electron beam excitation of a luminescent material. https://www.leica-microsystems.com//science-lab/a-word-on-cathodoluminescence/ Sun, 03 Feb 2013 23:00:00 +0000 Cathy Johnson https://www.leica-microsystems.com/7988 Confocal Microscopy Super-Resolution True confocal microscope systems feature single-point illumination and single-point detection. The method is called "optical sectioning" since the generated image contains only information from the focal plane. The serial detection offers highly efficient and low-noise sensors for signal conversion. Although the nonparallel detection is not conducive to high-speed imaging, modern scanning concepts allow frame rates above 400 frames per second at reasonable noise levels. This is by far enough for most applications, including the monitoring of fast ion-transport phenomena in living material. https://www.leica-microsystems.com//science-lab/gates-open-for-improved-confocal-fluorescence-and-super-resolution-sted/ Wed, 23 Jan 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/5734 EM Sample Preparation The University of Wollongong has a diverse range of materials research programs that includes metallurgy for mining, manufacturing, steel making and transport; polymers for solar cells, energy storage and bionic implants; and superconducting and electronic materials for commercialization, energy storage, telecommunications and medical applications. Electron microscopy is an integral part of this research, due to the chemical and structural information that can be provided down to the atomic scale. https://www.leica-microsystems.com//science-lab/university-of-wollongong-electron-microscopy-centre/ Thu, 17 Jan 2013 23:00:00 +0000 Darren Attard, Tony Romeo https://www.leica-microsystems.com/6656 Live-Cell Imaging Quantitative Imaging In eukaryotic cells Ca2+ is one of the most widespread second messengers used in signal transduction pathways. Intracellular levels of Ca2+ are usually kept low, as Ca2+ often forms insoluble complexes with phosphorylated and carboxylated compounds. Typically cytosolic Ca2+ concentrations are in the range of 100 nM. In response to stimuli Ca2+ may either be released from external medium or internal stores to raise the Ca2+ concentration. https://www.leica-microsystems.com//science-lab/widefield-calcium-imaging-with-calcium-indicator-fura2/ Sun, 13 Jan 2013 23:00:00 +0000 Gabriele Burger, Simone Diehl https://www.leica-microsystems.com/5728 EM Sample Preparation The Electron Microscopy (EM) laboratory in the School of Biological and Biomedical Sciences at Durham University is an integral part of a wider facility spanning a range of advanced imaging tools (laser scanning and spinning disc confocal microscopes, TIRF microscopy and live cell imaging) as well as an ultra high resolution (<0.5nm) field emission in-lens scanning electron microscope (Hitachi S5200) and a routine transmission EM with high tilt stage for tomography (Hitachi H7600). https://www.leica-microsystems.com//science-lab/biological-electron-microscopy/ Thu, 10 Jan 2013 23:00:00 +0000 Dr. Martin W. Goldberg https://www.leica-microsystems.com/7903 Super-Resolution At the Max Planck Institute for Biology of Ageing in Cologne founded in 2008, three departments are working on the molecular, physiological and evolutionary mechanisms of the ageing of cells, tissue and organisms. The long-term aim of the research is to help people age more healthily. Here, Dr. Christian Kukat from the department of Professor Nils-Göran Larsson is examining the role played by mitochondria in the ageing process. In close collaboration with Dr. Christian Wurm from the research group of Professor Stefan Jakobs at the Max Planck Institute for Biophysical Chemistry in Göttingen and researchers at the University of Gothenburg, Sweden, they harnessed super-resolution STED microscopy to find out new and surprising facts about the mitochondrial nucleoid. https://www.leica-microsystems.com//science-lab/mitochondrial-dna-molecules-are-packaged-individually/ Sun, 06 Jan 2013 23:00:00 +0000 Dr. Matthias Schauen, Dr. Christian Kukat https://www.leica-microsystems.com/7865 Basics in Microscopy Stereo Microscopy One hundred years ago, in 1913, the Optische Werke Ernst Leitz in Wetzlar, predecessor of Leica Microsystems CMS GmbH, made two inventions that were to blaze the trail for modern microscopy: the binocular tube and the integrating stage for quantitative microscopy. https://www.leica-microsystems.com//science-lab/100-years-of-binoculars-and-quantitative-microscopy/ Wed, 02 Jan 2013 23:00:00 +0000 Rolf Beck https://www.leica-microsystems.com/15675 Light Sheet Microscopy Light Sheet Fluorescence Microscopy (LISH-M) is a true fluorescence optical sectioning technique, first described by Heinrich Siedentopf in 1902 under the name of Ultramicroscopy. Light sheet microscopy utilises a plane of light to optically section samples. This allows deep imaging within transparent tissues and whole organisms. This book chapter will provide the reader with a comprehensive view on this emerging technology. https://www.leica-microsystems.com//science-lab/light-sheet-fluorescence-microscopy-beyond-the-flatlands/ Thu, 20 Dec 2012 14:05:00 +0000 https://www.leica-microsystems.com/7972 EM Sample Preparation One of the uses of the Scanning Electron Microscope (SEM) is in the study of surface morphology in biological applications which requires the preservation of the surface details of a specimen. Samples for Electron Microscopy (EM) imaging need to be dried in order to be compatible with the vacuum in the microscope. The presence of water molecules will disturb the vacuum and with it the imaging. https://www.leica-microsystems.com//science-lab/brief-introduction-to-critical-point-drying/ Mon, 10 Dec 2012 23:00:00 +0000 Dr. Ruwin Pandithage https://www.leica-microsystems.com/5723 EM Sample Preparation Atomic force microscopy (AFM) is a powerful tool in the suite of nanoscale characterization techniques that provides a variety of information including topography, mechanical properties, and electrical properties with nanoscale lateral and sub-nanometer vertical resolution. Cryoultramicrotomy is an essential tool for effective polymer sample preparation for atomic force microscopy (AFM) in order to get rid of the polymer skin from processing and to ensure a smooth surface for analysis. https://www.leica-microsystems.com//science-lab/atomic-force-microscopy-study-of-a-stretched-impact-copolymer/ Mon, 10 Dec 2012 23:00:00 +0000 Dr. Dalia G. Yablon, Jean Grabowski, Andy Haishung Tsou https://www.leica-microsystems.com/5738 EM Sample Preparation We have used cultured UMR106-01 osteoblastic cells to investigate the process of bone mineralization. UMR106-01 cells as well as primary calvarial bone cells assembly spherical extracellular supramolecular protein-lipid complexes, termed biomineralization foci (BMF), in which the first crystals of hydroxyapatite mineral are deposited (Midura et al., 2004; Wang et al., 2004). A major difference between these culture models is the speed with which mineralization occurs, ranging from 12–16 days after plating for primary osteoblastic cells to 88 h for UMR106-01 cells. https://www.leica-microsystems.com//science-lab/dry-ultrathin-sectioning-combined-with-high-pressure-freezing/ Mon, 10 Dec 2012 23:00:00 +0000 Ph.D. Jeff P. Gorski, M.Sc. Nichole T. Huffman, Thérèse Hillman-Marti, Daniel Studer https://www.leica-microsystems.com/7867 Super-Resolution The development and maintenance of polarized epithelial cells requires the establishment of complicated subcellular machinery. We studied the role of post-translational tubulin modifications within this process. At first, the distribution of detyrosinated microtubules was assessed in MDCK cells via immunofluorescence microscopy. No preferential accumulation of tyrosinated or detyrosinated microtubules could be detected at the apical or basal cell poles in epithelial cell cysts. However, during monolayer formation, the quantities of detyrosinated tubulin increased significantly over time. https://www.leica-microsystems.com//science-lab/tubulin-modifications-affect-monolayer-formation-and-apical-trafficking-in-epithelial-cells/ Tue, 04 Dec 2012 23:00:00 +0000 Dr. Sabrina Zink, Prof. Ralf Jacob https://www.leica-microsystems.com/7855 Quantitative Imaging Confocal Microscopy Live-Cell Imaging Fluorescence Microscopy Imaging techniques are indispensable in many fields of life sciences today. With state-of-the-art optics and metrology, they provide hundreds of gigabytes of still images and videos. Correspondingly, there is a growing need for complex software solutions to ensure that the amounts of generated data can be automatically managed, processed and analyzed – and shared online with a large group of users. The combination of individual open-source software projects is proving especially useful for solving such complex image analysis problems. https://www.leica-microsystems.com//science-lab/integrative-open-source-software-for-image-analysis-in-biology/ Tue, 27 Nov 2012 23:00:00 +0000 Christian Dietz, Martin Horn, Prof. Dr. Michael Berthold https://www.leica-microsystems.com/7817 Laser Microdissection Arbuscular mycorrhizal (AM) fungi are an essential feature of the biology and ecology of most terrestrial plants and, as biofertilizers, AM fungi are an emerging issue in many projects focused on low-input agriculture practices. The identification of the events that lead to the formation of an AM, including the mechanisms involved in nutrient transfer, will be a challenging objective for a better exploitation of AMs in agricultural programs. https://www.leica-microsystems.com//science-lab/quantitative-one-step-protocol-to-detect-transcripts-in-laser-microdissected-samples/ Wed, 21 Nov 2012 23:00:00 +0000 PhD Valentina Fiorilli, Marco Giovannetti, PhD Raffaella Maria Balestrini https://www.leica-microsystems.com/7797 Basics in Microscopy Stereo Microscopy Widefield Microscopy Modern image analysis systems perform highly sophisticated image processing functions on images from an automated microscope and digital camera. 50 years ago, the first image analysis system was analogue, based on a video camera and the area measurements could be read from a meter. Nevertheless, it marked the beginning of automation in this field. https://www.leica-microsystems.com//science-lab/50-years-of-image-analysis/ Sun, 18 Nov 2012 23:00:00 +0000 Geoff Jenkinson https://www.leica-microsystems.com/7793 Ergonomics Stereo Microscopy Nobody is the same. Whether large or small, well- or slightly built, left- or right-handed, everyone has different demands of the tool they work with for many hours a day. This is particularly true of laboratory workstations, where routine tasks such as pipetting or microscope or microtome work require a static posture. https://www.leica-microsystems.com//science-lab/body-in-balance/ Thu, 15 Nov 2012 23:00:00 +0000 John Ludescher https://www.leica-microsystems.com/7275 Laser Microdissection The physiological process of aging is an elementary part of life. The processes responsible for the aging process and the effect they have is one of the key focuses of biomedical research. Dr. Yvonne Begus-Nahrmann researches molecular mechanisms in stem cells and their role in aging and tumor development at the Göttingen University Medical Center. Signal proteins such as p53 and p21 appear to play an important role in this context. To investigate these proteins in specific cell types, the cells are isolated from organ tissue with the aid of laser microdissection. https://www.leica-microsystems.com//science-lab/stem-cells-in-the-aging-process-the-signal-protein-p53-as-guardian-of-the-genome/ Thu, 08 Nov 2012 23:00:00 +0000 M.Sc., Cornelia Gilbrich-Wille, Dr. Yvonne Begus-Nahrmann https://www.leica-microsystems.com/7735 Super-Resolution Widefield Microscopy The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. With a mixture of engaging talks by key experts in the field of super-resolution microscopy and stimulating discussion sessions, the meeting proved as popular as last year’s event, attracting a wide range of scientists interested in both confocal and widefield super-resolution and sample preparation techniques. https://www.leica-microsystems.com//science-lab/abstracts-of-the-2nd-european-super-resolution-user-club-meeting/ Thu, 01 Nov 2012 23:00:00 +0000 Prof. Hjalmar Brismar, Prof. Dr. Dr. h.c. Stefan Hell, Prof. Christian Eggeling, Dr. Anna Szymborska, Prof. Alberto Diaspro, Ph.D. Eric Hosy, Ph.D. Giuseppe Vicidomini, Hans van der Voort https://www.leica-microsystems.com/5222 Neuroscience Optogenetics is a technique that allows light-controlled responses of transfected cells. The cells are genetically modified by introduction of genes that code for light-induced channels or ion pumps. The term optogenetics denotes the light control feature introduced by genetic engineering. https://www.leica-microsystems.com//science-lab/optogenetics/ Mon, 29 Oct 2012 12:51:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/6828 Digital Microscopy Quality Assurance Digital microscopy offers clear advantages for a large number of industrial quality inspections, particularly for surface analysis. Here, you can find some videos that show examples of application for digital microscopy. https://www.leica-microsystems.com//science-lab/digital-microscopy/ Tue, 16 Oct 2012 22:00:00 +0000 Georg Schlaffer https://www.leica-microsystems.com/7364 Confocal Microscopy Live-Cell Imaging The Royal Swedish Academy of Sciences has decided to award the Nobel Prize in Chemistry for 2012 to Robert J. Lefkowitz and Brian K. Kobilka for studies of G-protein–coupled receptors. https://www.leica-microsystems.com//science-lab/nobel-prize-2012-in-chemistry-for-studies-of-g-protein-coupled-receptors/ Thu, 11 Oct 2012 22:00:00 +0000 Nobel Media AB 2012 https://www.leica-microsystems.com/7290 Confocal Microscopy Live-Cell Imaging The Nobel Prize recognizes two scientists who discovered that mature, specialised cells can be reprogrammed to become immature cells capable of developing into all tissues of the body. Their findings have revolutionised our understanding of how cells and organisms develop. https://www.leica-microsystems.com//science-lab/nobel-prize-2012-in-physiology-or-medicine-for-stem-cell-research/ Tue, 09 Oct 2012 22:00:00 +0000 Nobel Media AB 2012 https://www.leica-microsystems.com/7246 Multiphoton Microscopy Basics, history, and applications of multiphoton microscopy. Honouring Goeppert-Mayer’s prediction of simultaneous two-photon absorption by an atom or molecule reported in the 1930s in her PhD dissertation thesis, we can state that multiphoton excitation (MPE) microscopy, more frequently identified with two-photon excitation (2PE) fluorescence microscopy, is a key microscopy method in many areas from medicine to biology, from biophysics to materials science. https://www.leica-microsystems.com//science-lab/developments-in-multiphoton-excitation-microscopy/ Sun, 07 Oct 2012 22:00:00 +0000 Prof. Alberto Diaspro https://www.leica-microsystems.com/6773 Laser Microdissection Phasic activation of the dopamine (DA) midbrain system in response to unexpected reward or novelty is critical for adaptive behavioral strategies. This activation of DA midbrain neurons occurs via a synaptically triggered switch from low-frequency background spiking to transient high-frequency burst firing. We found that, in medial DA neurons of the substantia nigra (SN), activity of ATP-sensitive potassium (K-ATP) channels enabled NMDA-mediated bursting in vitro as well as spontaneous in vivo burst firing in anesthetized mice. https://www.leica-microsystems.com//science-lab/k-atp-channels-in-dopamine-substantia-nigra-neurons-control-bursting-and-novelty-induced-exploration/ Thu, 04 Oct 2012 22:00:00 +0000 Julia Schiemann, Prof. Birgit Liss, Dr. Gaby Schneider, Ph.D. Kareem A. Zaghloul, Dr. Peter J. Magill, Prof. Susumu Seino, Markus Bingmer, Dr. Verena Klose, Dr. Falk Schlaudraff, Prof. Jochen Röper https://www.leica-microsystems.com/6742 Laser Microdissection Due to its instability, RNA is generally more difficult to work with than DNA. RNA does not have the stability of the DNA double helix. To obtain the best results when dealing with RNA it is essential to start out with high-quality material and to carry out particularly careful quality control before and after processing. The so-called RIN (RNA Integrity Number) is an indicator of the quality of RNA. On a scale of 1–10, a RIN value of 1 indicates that the RNA is completely degraded and a RIN value of 10 that the RNA is fully intact. The higher the RIN number, the better the RNA quality. Wherever possible, material with high RIN numbers should always be used. https://www.leica-microsystems.com//science-lab/comparison-of-the-rna-quality-of-native-tissue-samples-with-tissue-samples-after-uv-laser-microdissection/ Sun, 30 Sep 2012 22:00:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/5141 Digital Microscopy Quality Assurance There are digital cameras, digital TV sets, digital picture frames, digital schools on the internet. Cryptologists design digital signatures, communication researchers speak of digital identity. Digital may be an overused buzzword, but digital technology has undeniably revolutionized our world ever since the invention of the computer and will continue to do so in future. https://www.leica-microsystems.com//science-lab/trends-in-microscopy/ Wed, 19 Sep 2012 22:00:00 +0000 Daniel Goeggel, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/6596 Digital Microscopy Quality Assurance One of the main features of a digital microscope is the speed and ease with which it enables surface models to be created of macroscopic and microscopic structures. In a qualitative evaluation, these provide a better understanding and a more detailed documentation of the specimen. In addition, quantification of the surface provides valuable information about the composition of the surface and its wear. Which specimens are suitable for use with a Leica digital microscope, and what are the limitations of the method used? https://www.leica-microsystems.com//science-lab/3d-visualization-of-surface-structures/ Wed, 19 Sep 2012 22:00:00 +0000 Daniel Goeggel, Georg Schlaffer https://www.leica-microsystems.com/6792 Live-Cell Imaging Stereo Microscopy In exploring how embryos take shape, John Wallingford has identified a key pathway involved in vertebrate development – and human disease. https://www.leica-microsystems.com//science-lab/taking-the-long-view/ Mon, 10 Sep 2012 22:00:00 +0000 https://www.leica-microsystems.com/6706 Forensics Stereo Microscopy Many people know Germany’s most famous forensic biologist, Dr. Mark Benecke from Cologne, from TV documentaries showing how crimes are solved. Benecke is a welcome guest on talk shows on topics such as forensic trace analysis, murder or the depths of the human psyche in general. He also enjoys an excellent international reputation. However, Mark Benecke’s normal working day bears little resemblance to the scenes shown in TV crime drama series. https://www.leica-microsystems.com//science-lab/even-insect-fragments-throw-light-on-how-crimes-are-committed-how-forensic-biologist-dr-mark-benecke-gains-insights/ Sun, 02 Sep 2012 22:00:00 +0000 M. A. Janika Wiesner, Dipl. oec.-troph. Anja Schué, Dr. Mark Benecke https://www.leica-microsystems.com/6673 Basics in Microscopy The traditional light source for fluorescence excitation is a fluorescence lamp housing with mercury burner. A prerequisite for achieving bright and homogeneous excitation is the correct centering and alignment of the bulb inside the housing. This video tutorial presents an easy-to-copy way to align the mercury bulb in a fluorescence lamp housing. https://www.leica-microsystems.com//science-lab/video-tutorial-how-to-align-the-bulb-of-a-fluorescence-lamp-housing/ Sun, 19 Aug 2012 22:00:00 +0000 Beate Braun, Dr. Werner Wittke https://www.leica-microsystems.com/6675 Basics in Microscopy When applying fluorescence microscopy in biological applications, a lamp housing with mercury burner is the most common light source. This video tutorial shows how to change the bulb of a traditional fluorescence lamp housing. https://www.leica-microsystems.com//science-lab/video-tutorial-how-to-change-the-bulb-of-a-fluorescence-lamp-housing/ Sun, 19 Aug 2012 22:00:00 +0000 Beate Braun, Dr. Werner Wittke https://www.leica-microsystems.com/6634 Forensics Stereo Microscopy “We have to take your fingerprints.” This sentence is spoken in nearly every TV crime drama to a suspect sitting in the interrogation room. But what exactly is it that makes a fingerprint so valuable for detectives in real life? How do fingerprint experts, known as dactyloscopists, perform their jobs? https://www.leica-microsystems.com//science-lab/how-a-fingerprint-is-traced-to-the-person-who-made-it-interactive-microscope-system-facilitates-dactyloscopist-training/ Sun, 12 Aug 2012 22:00:00 +0000 M. A. Janika Wiesner, Dieter Kauer https://www.leica-microsystems.com/6600 Live-Cell Imaging Confocal Microscopy Quantitative Imaging Many biological samples exhibit autofluorescence. Its often broad spectra can interfere with fluorescent labeling strategies. This application letter demonstrates how autofluorescence can serve as an intrinsic contrast in fluorescence lifetime imaging microscopy (FLIM) resulting in multi-color image stacks. https://www.leica-microsystems.com//science-lab/label-free-flim/ Thu, 09 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/6602 Live-Cell Imaging Confocal Microscopy Quantitative Imaging FLIM combines lifetime measurements with imaging: lifetimes obtained for each image pixel are color-coded to produce additional image contrast. Thus, FLIM delivers information about the spatial distribution of a fluorescent molecule together with information about its biochemical status or nano-environment. A typical application of FLIM is FLIM-FRET. FRET is a well-established technique to study molecular interactions. It scrutinizes protein binding and estimates intermolecular distances on an Angström scale as well. https://www.leica-microsystems.com//science-lab/fret-with-flim/ Tue, 07 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/6609 Confocal Microscopy Quantitative Imaging Fluorescence correlation spectroscopy (FCS) measures fluctuations of fluorescence intensity in a sub-femtolitre volume to detect such parameters as the diffusion time, number of molecules or dark states of fluorescently labeled molecules. The technique was independently developed by Watt Webb and Rudolf Rigler during the early 1970s. https://www.leica-microsystems.com//science-lab/fluorescence-correlation-spectroscopy/ Thu, 02 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/6499 Fluorescence Microscopy Live-Cell Imaging Widefield Microscopy This four-part series of articles published in Microscopy and Analysis covers the factors to consider in choosing a camera among CCD, EMCCD, and scientific-grade CMOS camera technologies for biological imaging applications. The differences among the sensor architectures and the impact of parameters such as pixel size, noise, and QE on signal-to-noise performance, image quality, and Nyquist sampling are considered. https://www.leica-microsystems.com//science-lab/digital-camera-technologies-for-scientific-bio-imaging/ Wed, 18 Jul 2012 22:00:00 +0000 https://www.leica-microsystems.com/5901 Coherent Raman Scattering (CRS) Coherent anti-Stokes Raman scattering (CARS) microscopy is a technique that generates images based on the vibrational signatures of molecules. This imaging methods does not require labeling, yet molecular specific information can be obtained from a range of important bio-molecular compounds. https://www.leica-microsystems.com//science-lab/cars-microscopy-imaging-characteristic-vibrational-contrast-of-molecules/ Mon, 16 Jul 2012 22:00:00 +0000 Ph.D. Eric Olaf Potma https://www.leica-microsystems.com/6497 Widefield Microscopy Super-Resolution Great advancements in biology have been possible by using fluorescence microscopy. So far, the resolution of the images was limited due to physical constraints. In the past couple of years, new methods evolved circumventing these limitations and bringing fluorescence microscopy to a new level of resolution, boosting the possibilities in science with fluorescence microscopes. https://www.leica-microsystems.com//science-lab/widefield-super-resolution-with-gsdim/ Wed, 11 Jul 2012 22:00:00 +0000 Dr. Marko Lampe https://www.leica-microsystems.com/6474 Basics in Microscopy Clean microscope optics are essential for obtaining good microscope images. If they are dirty, the microscope should be cleaned to avoid a loss of quality. If you decide to do this yourself, you should be extremely careful not to damage the sensitive microscope optics. https://www.leica-microsystems.com//science-lab/how-to-clean-microscope-optics/ Sun, 08 Jul 2012 22:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/6417 Widefield Microscopy Fluorescence Microscopy Live-Cell Imaging Fluorescence microscopy is a modern and steadily evolving tool to bring light to current cell biological questions. With the help of fluorescent proteins or dyes it is possible to make discrete cellular components visible in a highly specific manner. A prerequisite for these kinds of investigations is a powerful fluorescence microscope. One special aim is the three-dimensional illustration of a structure to get an impression of full plasticity. This poses a certain problem for the experimenter using a classical light microscope. https://www.leica-microsystems.com//science-lab/deconvolution/ Wed, 04 Jul 2012 22:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/6415 Confocal Microscopy The perfect light source for confocal microscopes in biomedical applications has sufficient intensity, tunable color and is pulsed for use in lifetime fluorescence. Furthermore, it should offer means to avoid reflection of excitation light, and the coupling into the beam path must be efficient and homogeneous throughout the full visible spectrum. Such a source has been invented and implemented: the white light laser in combination with acousto-optical beam splitting. https://www.leica-microsystems.com//science-lab/white-light-laser/ Tue, 26 Jun 2012 22:00:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Lioba Kuschel https://www.leica-microsystems.com/6409 Stereo Microscopy Every ant has a story to tell. And what better person to tell it than Brian Fisher? Fisher is an entomologist at the California Academy of Sciences and Project Manager of AntWeb.org. The internet portal illustrates the diversity of ants by providing information and high quality color images of many of the approximately 10,000 known species of ants. https://www.leica-microsystems.com//science-lab/antweb-documents-the-world-of-ants/ Mon, 18 Jun 2012 22:00:00 +0000 Dr. Brian L. Fisher https://www.leica-microsystems.com/6345 Fluorescence Microscopy Widefield Microscopy A basic principle in fluorescence microscopy is the highly specific visualization of cellular components with the help of a fluorescing agent. This can be a fluorescing protein – for example GFP – genetically linked to the protein of interest. If cloning is impossible – for instance in histologic samples – it is required to use other techniques like immunofluorescence staining to visualize the protein of interest. https://www.leica-microsystems.com//science-lab/fluorescent-dyes/ Sun, 10 Jun 2012 22:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/10886 Live-Cell Imaging EM Sample Preparation CLEM Despite the lipolysis–lipogenesis cycle being a fundamental process in adipocyte biology, very little is known about the morphological changes that occur during this process. The remodeling of lipid droplets to form micro lipid droplets (mLDs) is a striking feature of lipolysis in adipocytes, but once lipolysis ceases, the cell must regain its basal morphology. https://www.leica-microsystems.com//science-lab/postlipolytic-insulin-dependent-remodeling-of-micro-lipid-droplets-in-adipocytes/ Tue, 15 May 2012 14:14:00 +0000 https://www.leica-microsystems.com/5995 Stereo Microscopy Quality Assurance Exotic fruits and sun-kissed vegetables – we have long been accustomed to a huge selection of culinary delicacies that are available fresh in stores on a daily basis. Sometimes, however, these goods flown in from far away carry along unwanted passengers: pests, fungi, or viruses, which cannot be seen with the naked eye. https://www.leica-microsystems.com//science-lab/stereo-microscopes-in-the-eus-plant-inspection/ Sun, 06 May 2012 22:00:00 +0000 Claudia Moch https://www.leica-microsystems.com/5988 Basics in Microscopy The optical microscope has been a standard tool in life science as well as material science for more than one and a half centuries now. To use this tool economically and effectively, it helps a lot to understand the basics of optics, especially of those essential components which are part of every microscope. https://www.leica-microsystems.com//science-lab/optical-microscopes-some-basics/ Wed, 02 May 2012 22:00:00 +0000 Helmut Rühl https://www.leica-microsystems.com/5945 TIRF Microscopy The main feature of total internal reflection fluorescence (TIRF) microscopy is the employment of an evanescent wave for the excitation of fluorophores instead of using direct light. A property of the evanescent wave, which arises from the glass/water or glass/specimen interface, is that its propagation in z-direction gradually degrades, limiting its penetration depth into the specimen to some hundred nanometers. https://www.leica-microsystems.com//science-lab/controlling-the-tirf-penetration-depth-is-mandatory-for-reproducible-results/ Tue, 24 Apr 2012 22:00:00 +0000 Dr. Thomas Veitinger https://www.leica-microsystems.com/5947 Conservation Stereo Microscopy Through a period of two years an international research team has studied four almost identical paintings from the 16th century. The four paintings examined are alike yet different variations on a theme – Christ Driving the Traders from the Temple and were made with different purposes, answering to the demands of a booming 16th-century Antwerp art market. https://www.leica-microsystems.com//science-lab/four-paintings-magnified/ Sun, 22 Apr 2012 22:00:00 +0000 Anne Haack Christensen, Dr. Jørgen Wadum https://www.leica-microsystems.com/5929 Multiphoton Microscopy Confocal Microscopy In modern biomaterial design the generation of an environment mimicking some of the extracellular matrix features is envisaged to support molecular cross-talk between cells and scaffolds during tissue formation/remodeling. In bone substitutes chemical biomimesis has been particularly exploited; conversely, the relevance of pre-determined scaffold architecture for regenerated bone outputs is still unclear. https://www.leica-microsystems.com//science-lab/order-versus-disorder/ Thu, 19 Apr 2012 22:00:00 +0000 Paolo Giannoni, Paolo Bianco, Prof. Alberto Diaspro, Anna Tampieri, Ugo Valbusa, Giuseppe Firpo, Roberto Marotta, Monica Sandri, Paolo Bianchini, Rodolfo Quarto https://www.leica-microsystems.com/5899 Quantitative Imaging Seeing is believing – and measuring is knowing. Microscopes generate images that are not only used for illustration, but are also subject to quantification. More advanced techniques use illumination patterns (without image formation) or do not generate an image at all – but are still microscopical techniques. These F-techniques are becoming increasingly important in current biosciences. https://www.leica-microsystems.com//science-lab/quantitative-fluorescence/ Wed, 18 Apr 2012 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/5903 Neuroscience Confocal Microscopy Live-Cell Imaging Basics in Microscopy One of the crucial tasks of a research scientist is reporting and communicating about his work. This is vital for cutting edge research; it is crucial to gain insights from other experts, to get a discussion going, to improve, to be able to get some funding, to convince other colleagues that your research is good. Imagine that communication is not there. Then you will immediately appreciate its importance. There would be a lot of individuals all having a single piece of the puzzle in their hands, but nobody seeing the complete picture. https://www.leica-microsystems.com//science-lab/scientific-illustration-and-animation-for-research-scientists/ Sun, 15 Apr 2012 22:00:00 +0000 PhD Idoya Lahortiga, PhD Luk Cox https://www.leica-microsystems.com/5890 Fluorescence Microscopy Basics in Microscopy There are a lot of light-emitting processes occurring in nature. Luminescence is an umbrella term for those kinds of events where light emission is not the result of high temperatures. This article depicts the different forms of luminescences and goes into detail in the case of fluorescence. Relevant technical terms describing a fluorochrome, like quenching, bleaching or quantum yield, are explained in the second part of the article to give detailed insights into basic characteristics of fluorescent molecules. https://www.leica-microsystems.com//science-lab/basic-principles-of-luminescence/ Mon, 09 Apr 2012 22:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5831 Fluorescence Microscopy Widefield Microscopy The prospects of fluorescence microscopy changed dramatically with the discovery of fluorescent proteins in the 1950s. The starting point was the detection of the jellyfish Aequorea victoria green fluorescent protein (GFP) by Osamo Shimomura. Hundreds of GFP mutants later, the range of fluorescent proteins reaches from the blue to the red spectrum. https://www.leica-microsystems.com//science-lab/fluorescent-proteins-introduction-and-photo-spectral-characteristics/ Mon, 09 Apr 2012 22:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5106 Live-Cell Imaging The understanding of complex and fast cellular dynamics is an important step to get insight into biological processes. Therefore, today’s life science research more and more demands studying physiological events on the molecular level in real-time. https://www.leica-microsystems.com//science-lab/introduction-to-live-cell-imaging/ Tue, 03 Apr 2012 22:00:00 +0000 Dr. Thomas Veitinger https://www.leica-microsystems.com/5403 TIRF Microscopy Live-Cell Imaging Widefield Microscopy Total internal reflection fluorescence (TIRF) is a special technique in fluorescence microscopy developed by Daniel Axelrod at the University of Michigan, Ann Arbor in the early 1980s. TIRF microscopy delivers images with an outstandingly high axial resolution below 100 nm. This allows the observation of membrane-associated processes. https://www.leica-microsystems.com//science-lab/total-internal-reflection-fluorescence-tirf-microscopy/ Sun, 11 Mar 2012 23:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/5405 Live-Cell Imaging TIRF Microscopy Widefield Microscopy The special feature of TIRF microscopy is the employment of an evanescent field for fluorophore excitation. Unlike standard widefield fluorescence illumination procedures with arc lamps, LEDs or lasers, the evanescent field only penetrates the specimen by about 100 nm starting from the coverslip/medium interface. https://www.leica-microsystems.com//science-lab/applications-of-tirf-microscopy-in-life-science-research/ Sun, 11 Mar 2012 23:00:00 +0000 Wymke Ockenga, Dr. Thomas Veitinger https://www.leica-microsystems.com/5175 TIRF Microscopy Live-Cell Imaging Due to their special role in organ function and the exchange of biological components some body cells developed certain polarization characteristics. These are reflected in differences of their plasma membrane composition. The essential and fascinating task of polarized protein transport in epithelial cells is to get the right protein into the right membrane. https://www.leica-microsystems.com//science-lab/protein-transport-processes-at-the-apical-membrane-of-polarized-epithelial-cells/ Sun, 11 Mar 2012 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5391 Super-Resolution Natural killer (NK) cells are innate immune effectors that recognize and kill virally infected and tumorigenic cells. Central to their effector function is the formation of an immunological synapse with a target cell followed by directed secretion of cytolytic granules. https://www.leica-microsystems.com//science-lab/visualization-of-the-natural-killer-cell-immune-synapse-by-super-resolution-nanoscopy/ Thu, 08 Mar 2012 23:00:00 +0000 Ph.D. Emily M. Mace, Ph.D., MD Jordan S. Orange https://www.leica-microsystems.com/5331 Neuroscience Super-Resolution To explore the most intricate structures of the brain in order to decipher how it functions – Stefan Hell’s team of researchers at the Max Planck Institute for Biophysical Chemistry in Göttingen has made a significant step closer to this goal. Using the STED microscopy developed by Hell, the scientists have, for the first time, managed to record detailed live images inside the brain of a living mouse. https://www.leica-microsystems.com//science-lab/sharp-live-images-from-the-mouse-brain/ Sun, 04 Mar 2012 23:00:00 +0000 Dr. Sebastian Berning, Dr. Katrin Willig, Dr. Heinz Steffens, Dr. Payam Dibaj, Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/10210 Super-Resolution Natural killer (NK) cells are innate immune effectors that eliminate diseased and tumorigenic targets through the directed secretion of specialized secretory lysosomes, termed lytic granules. This directed secretion is triggered following the formation of an immunological synapse (IS), which is characterized by actin re-modeling and receptor organization at the interface between the NK cell and its susceptible target. https://www.leica-microsystems.com//science-lab/dual-channel-sted-nanoscopy-of-lytic-granules-on-actin-filaments-in-natural-killer-cells/ Thu, 01 Mar 2012 11:32:00 +0000 Ph.D. Emily M. Mace https://www.leica-microsystems.com/5321 Education Basics in Microscopy Bacteria are part of our world. There are countless numbers of them in the human body and they are completely harmless. But in people with a weak immune system or at the wrong place they can cause serious illness. Educational microscopes that pass through many hands are potential breeding grounds for germs. To solve this problem... https://www.leica-microsystems.com//science-lab/antimicrobial-coating-for-educational-microscopes/ Wed, 29 Feb 2012 23:00:00 +0000 M. A. Janika Wiesner, Christoph Fankhauser https://www.leica-microsystems.com/5108 Live-Cell Imaging Confocal Microscopy The understanding of complex and/or fast cellular dynamics is an important step for exploring biological processes. Therefore, today’s life science research is increasingly focusing on dynamic processes like cell migration, morphological changes of cells, organs or whole animals and physiological (e.g. changes of intracellular ion composition) events in living specimens in real time. https://www.leica-microsystems.com//science-lab/live-cell-imaging-techniques/ Thu, 23 Feb 2012 23:00:00 +0000 Dr. Thomas Veitinger, Dr. Zhongxiang Jiang https://www.leica-microsystems.com/5275 Confocal Microscopy Understanding how DNA lesions are optimally repaired is of functional significance, especially from the view of genome karyotype stability. https://www.leica-microsystems.com//science-lab/how-to-study-protein-recruitment-to-dna-lesions-by-a-combination-of-uv-laser-and-white-light-laser/ Wed, 22 Feb 2012 23:00:00 +0000 Assoc. Prof. Dr. Eva Bártová, Assoc. Prof. Dr. Stanislav Kozubek, Ph.D. Gabriela Sustáčková https://www.leica-microsystems.com/5268 Laser Microdissection Neuroscience Different areas of the Central Nervous System (CNS) display a specific and selective gene expression profile. Here, we used the Laser Microdissection system Leica LMD6500 to study region-specific mRNA expression in the adult mouse retina and hippocampus. https://www.leica-microsystems.com//science-lab/region-specific-gene-expression-in-adult-mouse-cns-tissues/ Tue, 21 Feb 2012 23:00:00 +0000 Andrea Messina, Mark Dunleavy, Ph.D. Paola Sgadò, Valentina Adami, Ph.D. Yuri Bozzi, Ph.D. Simona Casarosa https://www.leica-microsystems.com/5256 Fluorescence Microscopy Fluorescent proteins are the fundament of recent fluorescence microscopy and its modern applications. Their discovery and consequent development was one of the most exciting innovations for life sciences in the last century and the starting point of the deciphering of numberless natural phenomena. https://www.leica-microsystems.com//science-lab/fluorescent-proteins-from-the-beginnings-to-the-nobel-prize/ Thu, 16 Feb 2012 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5217 Fluorescence Microscopy Live-Cell Imaging Neuroscience TIRF Microscopy Confocal Microscopy Quantitative Imaging The need for data validation and accessibility has never been greater than it is today. We are inundated with information from a multitude of resources, but how can we easily evaluate the accuracy of that data? In the past, the peer review process provided this and was often run by publishers. https://www.leica-microsystems.com//science-lab/the-new-repository-on-the-block/ Mon, 06 Feb 2012 23:00:00 +0000 M.B.A. David N. Orloff, Ph.D. Janet Iwasa, Ph.D. Caroline Kane https://www.leica-microsystems.com/4658 Super-Resolution This video tutorial presents a reliable way to produce samples for super-resolution GSD imaging with a special focus on the mounting step. Stable and flat mounting of the coverslip increases the performance of the overall system, leading to an improved resolution of the GSD image. https://www.leica-microsystems.com//science-lab/video-tutorial-how-to-optimize-sample-preparation-for-gsd-microscopy/ Fri, 13 Jan 2012 10:09:00 +0000 Beate Braun, Dr. Marko Lampe https://www.leica-microsystems.com/5082 Confocal Microscopy Quantitative Imaging The characterization of substances at the single molecule level has become part of the standard repertoire of scientific research institutes. One of the most common methods is Fluorescence Correlation Spectroscopy (FCS), which can be used to examine the dynamics and concentration of fluorescent molecules in solution. https://www.leica-microsystems.com//science-lab/flcs-advances-in-fluorescence-correlation-spectroscopy/ Wed, 14 Dec 2011 23:00:00 +0000 Dr. Andreas Bülter, Dr. Andrea Bleckmann, Uwe Ortmann https://www.leica-microsystems.com/4758 Live-Cell Imaging Neuroscience Many fundamental functions of a cell strongly depend on delicate, but nevertheless dynamic balances of ions (e.g. calcium, magnesium), voltage potentials and pH between the cell’s cytosol and the surrounding extracellular space. Ratiometric imaging allows reliable estimations of ion concentrations and pH or voltage changes by measuring fluorophore emission shifts. https://www.leica-microsystems.com//science-lab/ratiometric-imaging/ Sun, 11 Dec 2011 23:00:00 +0000 Dr. Thomas Veitinger https://www.leica-microsystems.com/4487 Confocal Microscopy Fluorescence Microscopy usually employs incident light illumination. This requires a device that directs the light for illumination into the sample and transmits the light emitted by the sample to the detection system. In the past, various types of mirrors were the only option. Today, the acousto optical beam splitter serves best for the task. https://www.leica-microsystems.com//science-lab/beam-splitting/ Fri, 09 Dec 2011 10:23:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/4598 Confocal Microscopy To separate fractions of the emission for recording channels that reproduce the emission of individual fluorochromes, it is necessary to spatially disperse the emission spectrally. This is possible by employing dichroic mirrors or a genuine dispersive element like a prism or a grating. https://www.leica-microsystems.com//science-lab/spectral-imaging/ Fri, 09 Dec 2011 10:20:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/4465 Confocal Microscopy Fluorescence excitation needs specifically colored light. In confocal microscopy, multiline lasers or laser batteries are classically used. This requires devices that pick the requested lines fitting the currently employed fluorochromes. Intensity control is a second task that must be accomplished. https://www.leica-microsystems.com//science-lab/confocal-excitation/ Fri, 09 Dec 2011 07:59:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/4820 Super-Resolution Here we present an collection of videos telling the story of the invention of nanometer-scale microscopy by Stefan Hell and explaining the basic principle of STED microscopy. https://www.leica-microsystems.com//science-lab/videos-on-sted-microscopy-and-its-inventor-stefan-hell/ Thu, 08 Dec 2011 14:33:00 +0000 https://www.leica-microsystems.com/4849 Laser Microdissection Neuroscience How can we begin to understand the complexity of the human brain and the way it works? The same way we begin to understand a city: by making a map. In this talk, Allan Jones shows how his team is mapping which genes are turned on in each tiny region, and how it all connects up. https://www.leica-microsystems.com//science-lab/ted-talk-of-allan-jones-mapping-the-human-brain/ Fri, 02 Dec 2011 16:42:00 +0000 Ph.D. Allan Jones https://www.leica-microsystems.com/4867 Super-Resolution Stimulated Emission Depletion microscopy, or STED nanoscopy, is a technique that uses the non-linear de-excitation of fluorescent dyes to overcome the resolution limit imposed by diffraction encountered with standard confocal laser scanning microscopes and conventional far-field optical microscopes[1]. Compared to traditional confocal microscopy, STED offers exceptional improvements in resolution allowing visualization of cellular events at unprecedented levels. https://www.leica-microsystems.com//science-lab/subcellular-localization-of-akt-and-tubulin-using-super-resolution-microscopy/ Wed, 30 Nov 2011 23:00:00 +0000 Ph.D. David P. Chimento, Ph.D. Myriam Gastard, PhD Carl A. Ascoli https://www.leica-microsystems.com/4811 Confocal Microscopy Quantitative Imaging FRAP (Fluorescence recovery after photobleaching) can be used to study cellular protein dynamics: For visualization the protein of interest is fused to a fluorescent protein or a fluorescent dye. A region of interest (ROI) can be monitored applying a high amount of light to bleach the fluorescence within the ROI. The following illumination with low light conditions provides insight into the redistribution of molecules via recovery of fluorescence. https://www.leica-microsystems.com//science-lab/fluorescence-recovery-after-photobleaching-frap-and-its-offspring/ Wed, 23 Nov 2011 07:19:37 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/10159 Super-Resolution The advent of superresolution microscopy has opened up new research opportunities into dynamic processes at the nanoscale inside living biological specimens. This is particularly true for synapses, which are very small, highly dynamic, and embedded in brain tissue. Stimulated emission depletion (STED) microscopy, a recently developed laser-scanning technique, has been shown to be well suited for imaging living synapses in brain slices using yellow fluorescent protein as a single label. However, it would be highly desirable to be able to image presynaptic boutons and postsynaptic spines, which together form synapses, using two different fluorophores. https://www.leica-microsystems.com//science-lab/two-color-sted-microscopy-of-living-synapses-using-a-single-laser-beam-pair/ Sun, 20 Nov 2011 15:22:00 +0000 https://www.leica-microsystems.com/15671 Light Sheet Microscopy This chapter introduces the concept of light sheet microscopy along with practical advice on how to design and build such an instrument. Selective plane illumination microscopy is presented as an alternative to confocal microscopy due to several superior features such as high-speed full-frame acquisition, minimal phototoxicity, and multiview sample rotation. https://www.leica-microsystems.com//science-lab/quantitative-imaging-in-cell-biology-light-sheet-microscopy/ Sun, 20 Nov 2011 13:30:00 +0000 https://www.leica-microsystems.com/4713 Fluorescence Microscopy Here we present two review articles on fluorescent proteins and their biological applications. These first article reviews our current knowledge of blue, green, and red chromophore formation in permanently emitting FPs, photoactivatable FPs, and fluorescent timers. The second article focuses on novel monomeric RFPs and their application for studying gene expression, nuclear localization, and dynamics using advanced imaging. https://www.leica-microsystems.com//science-lab/modern-fluorescent-proteins-and-their-biological-applications/ Fri, 18 Nov 2011 12:50:00 +0000 https://www.leica-microsystems.com/6918 Super-Resolution The free online webinar on super-resolution presented by Leica Microsystems in association with Microscopy & Analysis took place on Tuesday, 15 November 2011. Register and view the webinar on demand. https://www.leica-microsystems.com//science-lab/applications-labeling-strategies-and-fluorophores-for-super-resolution/ Tue, 15 Nov 2011 15:52:00 +0000 Dr. Marko Lampe https://www.leica-microsystems.com/12640 Quality Assurance Interview with Dr. Michael Härtel, head of the materials testing laboratory at Continental Automotive GmbH, Powertrain Division, Limbach-Oberfrohna, Germany https://www.leica-microsystems.com//science-lab/technical-cleanliness-in-the-production-of-automotive-components/ Mon, 14 Nov 2011 17:06:00 +0000 Dipl. oec.-troph. Anja Schué, Dr. Michael Härtel https://www.leica-microsystems.com/4727 Neuroscience Live-Cell Imaging Especially in neuroscience, the physiology of ion channels has always been a major topic of interest. The development of the patch-clamp technique in the late 1970s has given electrophysiologists new prospects. It allows high-resolution current recordings not only of whole cells, but also of excised cellular patches. Even single-channel opening events can be investigated. However, with its complex technical, physical and biological background, the need for highly sensitive equipment and the huge amount of skills required of the experimenter, electrophysiology is still one of the most challenging methods in daily laboratory work. https://www.leica-microsystems.com//science-lab/the-patch-clamp-technique/ Wed, 09 Nov 2011 16:28:00 +0000 Dr. Sophie Veitinger https://www.leica-microsystems.com/4718 Neuroscience Neurobiology, the science of nerves and the brain, has mainly been driven forward in the last 200 years by microscopic investigations. The structures of cellular and subcellular structures, interaction and the three-dimensional assembly of neurons were made visible by various microscopy techniques. The optical microscope is also a necessary tool for visualizing micropipettes in electrophysiological measurements. Thirdly, many types of functional imaging are performed by means of optical microscopy. https://www.leica-microsystems.com//science-lab/neurobiology-and-microscopy/ Wed, 09 Nov 2011 13:07:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/4356 Confocal Microscopy Quantitative Imaging The Förster Resonance Energy Transfer (FRET) phenomenon offers techniques that allow studies of interactions in dimensions below the optical resolution limit. FRET describes the transfer of the energy from an excited state of a donor molecule to an acceptor molecule. Unlike absorption or emission of photons, FRET is a non-radiative energy exchange and consequently not a variation of light-matter interactions. https://www.leica-microsystems.com//science-lab/foerster-resonance-energy-transfer-fret/ Wed, 09 Nov 2011 09:45:00 +0000 Gabriele Burger https://www.leica-microsystems.com/4662 Coherent Raman Scattering (CRS) CARS overcomes the drawbacks of conventional staining methods by the intrinsic characteristics of the method. CARS does not require labeling because it is highly specific to molecular compounds which are based on vibrational contrast and chemical selectivity. The crucial advantage of this method is that the sample remains almost unaffected. With CARS, new samples that could not be stained due to unavailability of the appropriate dye are now accessible. https://www.leica-microsystems.com//science-lab/an-introduction-to-cars-microscopy/ Fri, 04 Nov 2011 14:36:46 +0000 Dr. Stefanie Degenhartt https://www.leica-microsystems.com/4438 Super-Resolution The nanoscopic technique GSDIM (ground state depletion microscopy followed by individual molecule return) provides a detailed image of the spatial arrangement of proteins and other biomolecules within the cell. There is now a first commercial system (the Leica SR GSD) on the market that is helping to make the GSDIM technique available to a wider group of users in research labs and imaging centers. https://www.leica-microsystems.com//science-lab/super-resolution-gsdim-microscopy/ Thu, 20 Oct 2011 22:00:00 +0000 PhD Tamara Straube, Alexandra Elli, Prof. Ralf Jacob https://www.leica-microsystems.com/10998 EM Sample Preparation Investigations of ultrastructural changes induced by viruses are often necessary to clearly identify viral diseases in plants. With conventional sample preparation for transmission electron microscopy (TEM) such investigations can take several days and are therefore not suited for a rapid diagnosis of plant virus diseases. Microwave fixation can be used to drastically reduce sample preparation time for TEM investigations with similar ultrastructural results as observed after conventionally sample preparation https://www.leica-microsystems.com//science-lab/microwave-assisted-rapid-diagnosis-of-plant-virus-diseases-by-transmission-electron-microscopy/ Fri, 14 Oct 2011 03:28:00 +0000 Prof. Bernd Zechmann, Prof. Günther Zellnig https://www.leica-microsystems.com/4392 Laser Microdissection Many of the discoveries that are now being made in cell division and differentiation, the relationships between single cells and cell organelles, treatments of cells with pharmaceutic substances, etc. would not be possible without live cell cultures. Allowing morphological and biochemical observations of single cells under different experimental conditions, they provide a unique source of information. https://www.leica-microsystems.com//science-lab/cell-cultures-and-laser-microdissection/ Fri, 07 Oct 2011 09:15:57 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/4384 Confocal Microscopy Confocal laser scanning microscopes are widely used to create highly resolved 3D images of cells, subcellular structures and even single molecules. Still, an increasing number of scientists are extending their focus of bio-research from single cell studies to entire organs and organisms, analyzing the complex interactions within whole animals. https://www.leica-microsystems.com//science-lab/mosaic-images/ Wed, 05 Oct 2011 12:34:00 +0000 Dr. Jan De Bock https://www.leica-microsystems.com/10212 Super-Resolution Accumulation of filamentous actin (F-actin) at the immunological synapse (IS) is a prerequisite for the cytotoxic function of natural killer (NK) cells. Subsequent to reorganization of the actin network, lytic granules polarize to the IS where their contents are secreted directly toward a target cell, providing critical access to host defense. https://www.leica-microsystems.com//science-lab/natural-killer-cell-lytic-granule-secretion-occurs-through-a-pervasive-actin-network-at-the-immune-synapse/ Thu, 15 Sep 2011 11:00:00 +0000 https://www.leica-microsystems.com/10165 Super-Resolution Neuroscience Live-Cell Imaging It is difficult to investigate the mechanisms that mediate long-term changes in synapse function because synapses are small and deeply embedded inside brain tissue. Although recent fluorescence nanoscopy techniques afford improved resolution, they have so far been restricted to dissociated cells or tissue surfaces. However, to study synapses under realistic conditions, one must image several cell layers deep inside more-intact, three-dimensional preparations that exhibit strong light scattering, such as brain slices or brains in vivo. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-of-actin-dynamics-in-synapses-deep-inside-living-brain-slices/ Wed, 07 Sep 2011 16:39:00 +0000 https://www.leica-microsystems.com/10866 CLEM Fluorescence Microscopy EM Sample Preparation Electron microscopy (EM) achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ visualization in cells and tissues. Here we introduce ‘"miniSOG"’ (for mini Singlet Oxygen Generator), a fluorescent flavoprotein engineered from Arabidopsis phototropin 2. https://www.leica-microsystems.com//science-lab/a-genetically-encoded-tag-for-correlated-light-and-electron-microscopy-of-intact-cells-tissues-and-organisms/ Sun, 04 Sep 2011 19:37:00 +0000 https://www.leica-microsystems.com/5122 Metallography Quality Assurance The examination of microstructure morphology plays a decisive role in materials science and failure analysis. There are many possibilities of visualizing the real structures of materials in the light microscope. The image samples shown in this article demonstrate the information potential of some of the techniques used. https://www.leica-microsystems.com//science-lab/metallography-with-color-and-contrast/ Tue, 30 Aug 2011 22:00:00 +0000 Ursula Christian, Prof. Dipl. Wirt. Ing. Norbert Jost https://www.leica-microsystems.com/5124 Forensics Stereo Microscopy ID cards, driving licenses, birth certificates, A-level exams – the potential for individual perpetrators or gangs to gain advantages by forging documents is vast. And the more sophisticated the security standards, the better equipped experts have to be in order to clearly differentiate between genuine documents and fakes. https://www.leica-microsystems.com//science-lab/genuine-or-fake/ Tue, 30 Aug 2011 22:00:00 +0000 Dipl. oec.-troph. Anja Schué, Martin Fischer https://www.leica-microsystems.com/4339 Conservation Stereo Microscopy “Germanic barbarians defeat super army!” That is the kind of news headline you might have seen in the year 9 AD about the victory of the Germanic tribes over three Roman legions under the command of Publius Quinctilius Varus. The Battle of the Teutoburg Forest is regarded as one of the most momentous battles of antiquity, and for a long time scientists have puzzled over where the fighting may have taken place. https://www.leica-microsystems.com//science-lab/where-the-germanic-forces-beat-the-romans/ Tue, 30 Aug 2011 22:00:00 +0000 Gisela Söger https://www.leica-microsystems.com/5118 Forensics Digital Microscopy Nobody wants counterfeit money in their wallet. Even counterfeiters want to get rid of their own creations as quickly as possible. However, experts work intensively to identify counterfeit money on behalf of the law. Martin Weber of the National Analysis Center of the German Bundesbank (Federal Bank) in Mainz is such an expert on forged banknotes. https://www.leica-microsystems.com//science-lab/collecting-funny-money/ Thu, 11 Aug 2011 22:00:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/4126 Confocal Microscopy Fluorescence Microscopy Fast True Confocal Scanning reduces photobleaching and increases the fluorescence yield at identical acquisition times. The long-lasting triplet state (or any other “dark state”) is less populated when the illumination is applied in shorter pulses at the same intensity. Consequently, more fluorochromes are available for the fluorescence process (brighter images) and fewer fluorochromes disintegrate from triplet states or excited triplet states (less bleaching). https://www.leica-microsystems.com//science-lab/brighter-fluorescence-by-resonant-scanning/ Wed, 03 Aug 2011 08:42:54 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/4122 Confocal Microscopy The obvious has been explored. These days, biologists strive to identify and analyze hidden and rare events. The task is tackled by automatically screening large numbers of objects – typically growing in multi-well plates – over a long period of time. When an interesting feature is identified (manually or by means of computed recognition), modern systems can automatically monitor and record these events at high resolution. https://www.leica-microsystems.com//science-lab/find-the-needle-in-the-haystack/ Wed, 03 Aug 2011 06:58:48 +0000 Simone Diehl https://www.leica-microsystems.com/11027 EM Sample Preparation Neuroscience This protocol describes how biological samples, like brain tissue, can be imaged in three dimensions using the focussed ion beam/scanning electron microscope (FIB/SEM). The samples are fixed with aldehydes, heavy metal stained using osmium tetroxide and uranyl acetate. They are then dehydrated with alcohol and infiltrated with resin, which is then hardened. https://www.leica-microsystems.com//science-lab/focussed-ion-beam-milling-and-scanning-electron-microscopy-of-brain-tissue/ Wed, 06 Jul 2011 16:17:00 +0000 https://www.leica-microsystems.com/3819 Laser Microdissection Laser microdissection is a highly selective process for preparing samples for DNA, RNA and protein analysis. It is a microscope-controlled manipulation technique for the precise separation of samples and tissue using a focused laser beam. https://www.leica-microsystems.com//science-lab/an-introduction-to-laser-microdissection/ Tue, 28 Jun 2011 15:39:57 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/3835 Laser Microdissection Laser microdissection and laser micromanipulation are suitable for gaining a differentiated insight into the function of genes and proteins, and are used for a wide range of applications in neurobiology and immunology as well as in the developmental and cell biology of animal and plant organisms. https://www.leica-microsystems.com//science-lab/applications-of-laser-microdissection/ Sun, 26 Jun 2011 22:00:00 +0000 M.Sc., Cornelia Gilbrich-Wille https://www.leica-microsystems.com/3772 Confocal Microscopy True confocal scanning microscopes (TCS) use a variable detection pinhole. Good optical sectioning tries to use just the inner core of the PSF. https://www.leica-microsystems.com//science-lab/pinhole-controls-optical-slicing/ Tue, 21 Jun 2011 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/3720 Confocal Microscopy Confocal microscopes are employed to optically slice comparably thick samples. https://www.leica-microsystems.com//science-lab/confocal-optical-section-thickness/ Mon, 20 Jun 2011 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/3768 Confocal Microscopy True confocal scanning microscopy (TCS) requires bright diffraction-limited illumination. https://www.leica-microsystems.com//science-lab/lasers-for-confocal/ Mon, 20 Jun 2011 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/10218 Super-Resolution Live-Cell Imaging We report superresolution fluorescence microscopy in an intact living organism, namely Caenorhabditis elegans nematodes expressing green fluorescent protein (GFP)-fusion proteins. We also superresolve, by stimulated emission depletion (STED) microscopy, living cultured cells, demonstrating that STED microscopy with GFP can be widely applied. https://www.leica-microsystems.com//science-lab/nanoscopy-in-a-living-multicellular-organism-expressing-gfp/ Wed, 15 Jun 2011 12:38:00 +0000 https://www.leica-microsystems.com/3660 Basics in Microscopy Widefield Microscopy Optical contrast methods give the potential to easily examine living and colorless specimens. Different microscopic techniques aim to change phase shifts caused by the interaction of light with the specimen into amplitude shifts that are visible to the human eye as differences in brightness. https://www.leica-microsystems.com//science-lab/optical-contrast-methods/ Thu, 09 Jun 2011 22:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/3662 Basics in Microscopy Widefield Microscopy Phase contrast is an optical contrast technique for making unstained phase objects (e.g. flat cells) visible under the light microscope. Cells that appear inconspicuous and transparent in brightfield can be viewed in high contrast and rich detail using phase contrast microscopy. https://www.leica-microsystems.com//science-lab/phase-contrast/ Wed, 08 Jun 2011 22:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/3664 Basics in Microscopy Widefield Microscopy Differential interference contrast (DIC) microscopy is a good alternative to brightfield microscopy for gaining proper images of unstained specimens that often only provide a weak image in brightfield. https://www.leica-microsystems.com//science-lab/differential-interference-contrast-dic/ Mon, 06 Jun 2011 22:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/3287 Basics in Microscopy Widefield Microscopy Hoffman modulation contrast has established itself as a standard for the observation of unstained, low-contrast biological specimens. The integration of the modulator in the beam path of themodern inverted microscopes allows a wide range of brightfield or phase objectives to be used, rather than a small selection of special objectives. https://www.leica-microsystems.com//science-lab/integrated-modulation-contrast-imc/ Tue, 31 May 2011 22:00:00 +0000 Dipl.-Phys. Bernard Kleine, Dr. Thomas Veitinger https://www.leica-microsystems.com/4182 Fluorescence Microscopy Basics in Microscopy Fluorescence is widely used in microscopy and an important tool for observing the distribution of specific molecules. Most molecules in cells do not fluoresce. They therefore have to be marked with fluorescing molecules called fluorochromes. https://www.leica-microsystems.com//science-lab/an-introduction-to-fluorescence/ Tue, 31 May 2011 13:29:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/4186 Basics in Microscopy Fluorescence Microscopy Molecules and atoms can exist in different quantum states. These states are dedicated to different energy levels; the quantum state with the lowest energy is called the ground state. Every state of greater energy is an excited state of the quantum mechanical system. Electrons can be excited by an exogenous energy source and switch to a higher energy level, changing the quantum state of the molecule or atom. The electrons are often referred to as being brought to a state of higher energy. https://www.leica-microsystems.com//science-lab/photo-effects-of-light/ Mon, 30 May 2011 15:13:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/4195 Basics in Microscopy Widefield Microscopy Polarization microscopy is routinely applied in material sciences and geology to identify minerals on the basis of characteristic refraction properties and colors. In biology, polarization microscopy is commonly used for identification or imaging of birefringent structures like crystals, or for imaging of cellulose in cell walls of plants and starch grains. https://www.leica-microsystems.com//science-lab/polarization-contrast/ Thu, 26 May 2011 14:08:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/9689 Neuroscience Confocal Microscopy Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 and have constituted a major breakthrough in the field of neuroscience. Since then, the technique was developed further and currently there are many different ways to prepare organotypic cultures. The method presented here was adapted from the one described by Stoppini et al. for the preparation of the slices and from Gogolla et al. for the staining procedure. https://www.leica-microsystems.com//science-lab/organotypic-cerebellar-cultures-apoptotic-challenges-and-detection/ Tue, 17 May 2011 17:50:00 +0000 https://www.leica-microsystems.com/2860 Confocal Microscopy “Confocal Microscopy” refers to a particular optical microscope that allows recording optical sections. Optical sectioning is achieved in a confocal system by illuminating and observing a single diffraction limited spot. https://www.leica-microsystems.com//science-lab/confocal-microscopy-optical-path/ Mon, 09 May 2011 12:36:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/10321 Quantitative Imaging FRET-based sensors for cyclic Adenosine Mono Phosphate (cAMP) have revolutionized the way in which this important intracellular messenger is studied. The currently prevailing sensors consist of the cAMP-binding protein Epac1, sandwiched between suitable donor- and acceptor fluorescent proteins (FPs). https://www.leica-microsystems.com//science-lab/a-mturquoise-based-camp-sensor-for-both-flim-and-ratiometric-read-out-has-improved-dynamic-range/ Fri, 29 Apr 2011 14:47:00 +0000 https://www.leica-microsystems.com/4188 Fluorescence Microscopy Basics in Microscopy Widefield Microscopy Fluorescence microscopy is a special form of light microscopy. It uses the ability of fluorochromes to emit light after being excited with light of a certain wavelength. Proteins of interest can be marked with such fluorochromes via antibody staining or tagging with fluorescent proteins. https://www.leica-microsystems.com//science-lab/fluorescence-in-microscopy/ Wed, 27 Apr 2011 15:47:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/3144 Stereo Microscopy An insight into the unknown world of the deep sea was given in an exhibition staged by the Senckenberg Society together with the Natural History Museum in Basel, Switzerland. Shown in Frankfurt am Main and Berlin, Germany, the "Deep Sea" exhibition was a huge public success in 2009. From May to September 2010, the exhibition has been shown in the Natural History Museum in London. Scientists of the Senckenberg Society work with stereomicroscopes and digital cameras both in the laboratory and on the research ships. https://www.leica-microsystems.com//science-lab/one-of-our-last-horizons/ Fri, 22 Apr 2011 22:00:00 +0000 Prof. Dr. Michael Türkay https://www.leica-microsystems.com/3862 Laser Microdissection Neuroscience Laser dissection is used in a large number of research fields, e.g. neurology, cancer research, plant analysis. Here, user report on the research results they have attained by using laser microdissection. https://www.leica-microsystems.com//science-lab/users-report-on-the-relevance-of-laser-microdissection-for-their-research-results/ Tue, 12 Apr 2011 22:00:00 +0000 https://www.leica-microsystems.com/3669 Coherent Raman Scattering (CRS) Multiphoton Microscopy In recent years, new molecular imaging techniques, such as coherent anti-Stokes Raman scattering microscopy (CARS), have been developed for rapid vibrational imaging of living cells. https://www.leica-microsystems.com//science-lab/good-vibrations/ Tue, 12 Apr 2011 22:00:00 +0000 Ph.D. Eric Olaf Potma, Dr. Bernd Sägmüller https://www.leica-microsystems.com/2994 Live-Cell Imaging TIRF Microscopy Application of TIRF microscopy (Total Internal Reflection Fluorescence) allows the visualization of structures at the apical surface of polarized epithelial cells that have been hidden in conventional fluorescence microscopy images. Hence, the approach reveals new insights into the composition of this characteristic cell pole that elucidate processes in apical protein trafficking. https://www.leica-microsystems.com//science-lab/tirf-microscopy-of-the-apical-membrane-of-polarized-epithelial-cells/ Tue, 12 Apr 2011 22:00:00 +0000 Dr. Christoph Greb, Prof. Ralf Jacob https://www.leica-microsystems.com/3000 Live-Cell Imaging Neuroscience A combination of widefield imaging techniques and image segmentation analysis enable researchers to map learning-induced functional changes in individual synapses throughout the hippocampus. https://www.leica-microsystems.com//science-lab/mapping-billions-of-synapses-with-microscopy-and-mathematics/ Tue, 12 Apr 2011 22:00:00 +0000 Dr. Christopher S. Rex, Allison Paradise https://www.leica-microsystems.com/3514 Confocal Microscopy Quantitative Imaging Although time correlated single photon counting (TCSPC) is the method of choice for fluorescence lifetime quantification, it requires dedicated instrumentation including a pulsed laser source, a photon counting card, and a fast detector. https://www.leica-microsystems.com//science-lab/choose-your-excitation-wavelength/ Tue, 12 Apr 2011 22:00:00 +0000 Ph.D. Corentin Spriet, Ph.D. Aymeric Leray, Dave Trinel, Franck Riquet, Laurent Héliot https://www.leica-microsystems.com/15669 Light Sheet Microscopy Light sheet fluorescence microscopy (LSFM) functions as a non-destructive microtome and microscope that uses a plane of light to optically section and view tissues with subcellular resolution. This method is well suited for imaging deep within transparent tissues or within whole organisms, and because tissues are exposed to only a thin plane of light, specimen photobleaching and phototoxicity are minimized compared to wide-field fluorescence, confocal, or multiphoton microscopy. https://www.leica-microsystems.com//science-lab/light-sheet-fluorescence-microscopy-a-review/ Sun, 20 Feb 2011 12:47:00 +0000 https://www.leica-microsystems.com/10323 Quantitative Imaging This study employed fluorescence lifetime spectroscopy to investigate the influence of ATP (adenosine triphosphate) on the fluorescence intensities of yellow and cyan fluorescent proteins (YFP/CFP) which are used as the basis for FRET (Förster resonance energy transfer) sensors. This work further elucidates the complex and heterogeneous fluorescence properties of CFP and its mutant H148D, as well as the sensitivity of CFP fluorescence to changes in solute conditions. https://www.leica-microsystems.com//science-lab/atp-changes-the-fluorescence-lifetime-of-cyan-fluorescent-protein-via-an-interaction-with-his148/ Fri, 05 Nov 2010 16:34:00 +0000 Eugenia Petoukhov, PhD James DeRose https://www.leica-microsystems.com/2470 Live-Cell Imaging Neuroscience Yet we are only just beginning to understand the complexities and functional differences of the sense of smell in mammals. Prof. Marc Spehr, head of the Department of Chemosensation at RWTH Aachen University since 2009, explains his findings on the neuronal mechanisms of olfactory perception and signal processing using the mouse model. He and his team are trying to find out how substances for social interaction are perceived and how this perception generates a specific type of behavior. https://www.leica-microsystems.com//science-lab/sniffing-out-the-secrets-of-social-behavior/ Mon, 01 Nov 2010 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, Prof. Dr. Marc Spehr https://www.leica-microsystems.com/2471 Neuroscience Super-Resolution Fully understanding the functionality and complexity of the human central nervous system remains one of the major open questions in modern science. Stimulated emission depletion microscopy (STED) can be the method to reveal biological nanostructures https://www.leica-microsystems.com//science-lab/the-missing-link-to-the-nanocosm-of-life/ Mon, 01 Nov 2010 23:00:00 +0000 https://www.leica-microsystems.com/2727 Neuroscience Super-Resolution Synapses are the switch-points in our brain for information transmission, learning and memory. News studies and developments of imaging techniques have provided new insights into the dynamics of glutamate receptors. The use of superresolution technologies is making an essential contribution to this research. https://www.leica-microsystems.com//science-lab/restless-receptors/ Mon, 01 Nov 2010 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, PhD Daniel Choquet https://www.leica-microsystems.com/2730 Multiphoton Microscopy Confocal Microscopy Developmental biology using Multiphoton microscopy with OPO. To gain new insight into the fundamental control of cell response to physical changes and to study the dynamics and roles of biological flow during the development of the zebrafish, Dr. Julien Vermot established his lab last year at the Institute of Genetics and Molecular and Cellular Biology (IGBMC) in Strasbourg, France. https://www.leica-microsystems.com//science-lab/deep-tissue-imaging/ Mon, 01 Nov 2010 23:00:00 +0000 PhD Julien Vermot, Dr. Andrea Mülter https://www.leica-microsystems.com/2731 Confocal Microscopy Multiphoton Microscopy Multiphoton microscopy with OPO: imaging with excitation wavelengths up to 1.300 nm. Because light scattering is dependent on the wavelength, better tissue penetration can be achieved by using longer excitation wavelengths. This is where excitation with infrared light, two-photon processes, and the OPO (optical parameter oscillator) can dramatically improve image quality. https://www.leica-microsystems.com//science-lab/what-is-an-opo/ Mon, 01 Nov 2010 23:00:00 +0000 Dr. Bernd Sägmüller, Dr. Andrea Mülter https://www.leica-microsystems.com/2732 Confocal Microscopy Multiphoton Microscopy Live-Cell Imaging Neuroscience Brain research using Confocal and Multiphoton Microscopy. Using imaging techniques such as confocal and two-photon microscopy, neuronal dendritic arborization of neurons and their synaptic interconnections can be visualized. https://www.leica-microsystems.com//science-lab/exploring-the-concert-of-neuronal-activities/ Mon, 01 Nov 2010 23:00:00 +0000 Ph.D. Randy Bruno, Ph.D. Myriam Gastard https://www.leica-microsystems.com/2743 Coherent Raman Scattering (CRS) Multiphoton Microscopy Confocal Microscopy The most important drawback of single-photon and multiphoton confocal microscopy is the need to label the specimen. CARS (Coherent Anti-Stokes Raman Spectroscopy) addresses this issue because it is non-toxic, non-destructive, and minimally invasive. https://www.leica-microsystems.com//science-lab/cars-and-confocal/ Mon, 01 Nov 2010 23:00:00 +0000 Dr. Stefanie Degenhartt, Ph.D. Vanessa Lurquin https://www.leica-microsystems.com/9687 Neuroscience We will demonstrate how to study the functional effects of introducing a point mutation in an ion channel. We study G protein-gated inwardly rectifying potassium (referred to as GIRK) channels, which are important for regulating the excitability of neurons. There are four different mammalian GIRK channel subunits (GIRK1-GIRK4) – we focus on GIRK2 because it forms a homotetramer. https://www.leica-microsystems.com//science-lab/mutagenesis-and-functional-analysis-of-ion-channels-heterologously-expressed-in-mammalian-cells/ Fri, 01 Oct 2010 16:35:00 +0000 https://www.leica-microsystems.com/10214 Super-Resolution Neuroscience Neurotransmitter release relies on the fusion of synaptic vesicles with the plasma membrane of synaptic boutons, which is followed by the recycling of vesicle components and formation of new vesicles. It is not yet clear whether upon fusion the vesicles persist as multimolecular patches in the plasma membrane, or whether they segregate into individual components. https://www.leica-microsystems.com//science-lab/the-fate-of-synaptic-vesicle-components-upon-fusion/ Fri, 01 Oct 2010 11:41:00 +0000 Dr. Felipe Opazo https://www.leica-microsystems.com/9478 Surgical Microscopy Before academic education for medical practitioners became the norm, barber-surgeons treated wounds and performed minor operations as members of the tradesmen’s guild as late as the end of the 16th century. The surgeon Wilhelm Fabry, born in Hilden in 1560, Germany, was not content with this classification of his profession, however. https://www.leica-microsystems.com//science-lab/wilhelm-fabry-surgeon-inventor-and-publicist/ Fri, 13 Aug 2010 19:06:00 +0000 M. A. Janika Wiesner, Dr. Wolfgang Antweiler https://www.leica-microsystems.com/9472 Surgical Microscopy In pars plana vitrectomy, three ocular incisions are normally made. Thanks to the superb optics and the unique illumination concept of the Leica M844 F40, Dr. Luca Cappuccini from Reggio Emilia Hospital in Italy can operate without one of the incisions for certain vitreoretinal procedures. This shortens the duration of surgery and speeds up eye recovery time. https://www.leica-microsystems.com//science-lab/state-of-the-art-microscope-enables-new-vitreoretinal-surgery-technique/ Fri, 13 Aug 2010 17:54:00 +0000 Dipl. oec.-troph. Anja Schué, Dr. Luca Cappuccini https://www.leica-microsystems.com/4411 Super-Resolution For centuries, cell biology has been based on light microscopy and at the same time been limited by its optical resolution. However, several new technologies have been developed recently that bypass this limit. https://www.leica-microsystems.com//science-lab/a-guide-to-super-resolution-fluorescence-microscopy/ Sun, 18 Jul 2010 22:00:00 +0000 https://www.leica-microsystems.com/5112 Stereo Microscopy Surgical Microscopy Ergonomics Quality Assurance Retina surgery demands experienced surgeons and precision technology. In vitreoretinal surgery, the surgeon operates with microscissors and forceps that are less than a millimeter thick. The Swiss company Alcon Grieshaber is one of the world’s leading specialists in handheld instruments for minimally invasive eye surgery. https://www.leica-microsystems.com//science-lab/precision-that-saves-eyesight/ Wed, 28 Apr 2010 22:00:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/5114 Stereo Microscopy Quality Assurance Only a few decades ago, the diagnosis ‘cataracts’ meant loss of vision in the near future. Today, cataract surgery is the most common operation worldwide. https://www.leica-microsystems.com//science-lab/intraocular-lenses/ Wed, 28 Apr 2010 22:00:00 +0000 https://www.leica-microsystems.com/5116 Metallography Quality Assurance Steel is a real all-rounder. However, each application requires a specifi c sort of steel grade. Without steel there would be no Olympic stadiums, wind energy plants, bridges, skyscrapers, trains, planes, cars, razor blades or knives for medical and home use – at least, not of the quality and design we know today. Buderus Edelstahl GmbH in Wetzlar, Germany is one of the world’s top producers of special steel. https://www.leica-microsystems.com//science-lab/steel-the-all-rounder-that-has-to-pass-many-tests/ Wed, 28 Apr 2010 22:00:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/2557 Basics in Microscopy Stereo Microscopy Widefield Microscopy In microscopy, depth of field is often seen as an empirical parameter. In practice it is determined by the correlation between numerical aperture, resolution and magnification. For the best possible visual impression, the adjustment facilities of modern microscopes produce an optimum balance between depth of field and resolution – two parameters which in theory are inversely correlated. https://www.leica-microsystems.com//science-lab/how-sharp-images-are-formed/ Wed, 28 Apr 2010 22:00:00 +0000 Ruedi Rottermann, Dipl. Geol. Peter Bauer https://www.leica-microsystems.com/2561 Multiphoton Microscopy Live-Cell Imaging Confocal Microscopy The comprehension of stem cell biology and its molecular basis is now acquiring paramount importance in cancer research. The need to look at a single, possibly living, cell makes fluorescence microscopy and confocal microscopy invaluable allies in the study of stem cells. https://www.leica-microsystems.com//science-lab/stem-cell-biology-in-cancer-research/ Thu, 22 Apr 2010 22:00:00 +0000 Laura Furia, Angelo Cicalese, Ilaria Rancati, Isabella Ponzanelli, Paolo Bianchini, Prof. Alberto Diaspro, Mario Faretta https://www.leica-microsystems.com/2718 Super-Resolution Scientists strive to understand the architecture of life. They want to learn how biological structures are arranged in respect to one another. Multicolor superresolution imaging allows fundamental questions to be addressed by far-field fluorescence microscopy in unprecedented detail. https://www.leica-microsystems.com//science-lab/confocal-nanoscopy-goes-multicolor/ Thu, 11 Feb 2010 10:06:00 +0000 PhD Jochen J. Sieber https://www.leica-microsystems.com/10831 CLEM EM Sample Preparation Microscopy is an essential tool for analysis of cellular structures and function. With the advent of new fluorescent probes and super-resolution light microscopy techniques, the study of dynamic processes in living cells has been greatly facilitated. Fluorescence light microscopy provides analytical, quantitative, and three-dimensional (3D) data with emphasis on analysis of live cells using fluorescent markers. https://www.leica-microsystems.com//science-lab/advanced-correlative-lightelectron-microscopy-current-methods-and-new-developments-using-tokuyasu-cryosections/ Sun, 20 Dec 2009 09:11:00 +0000 https://www.leica-microsystems.com/4362 Conservation Stereo Microscopy In the 2nd century BC, Baiae in the Gulf of Naples was a notorious bathing resort and spa for wealthy Romans. Today, part of the town is submerged under the sea and can be visited as an 80,000 square metre archaeological underwater park. The magnificent mosaics from the underwater ruins are analysed at the Istituto Superiore per la Conservazione e il Restauro (ISCR) of the Ministry for Cultural Heritage and Activities in Rome. https://www.leica-microsystems.com//science-lab/antique-underwater-treasures-endangered/ Mon, 14 Dec 2009 23:00:00 +0000 Gianfranco Priori https://www.leica-microsystems.com/4364 Stereo Microscopy Gemstones have fascinated people for thousands of years. Rulers and kings used to demonstrate their power and wealth with jewel-studded insignia. Although fine jewellery is still a status symbol of the rich, we now tend to treasure these wonders of nature more for our own pleasure in beauty and harmony. https://www.leica-microsystems.com//science-lab/exclusive-aesthetics-of-nature/ Mon, 14 Dec 2009 23:00:00 +0000 M.Sc. Michael Hügi https://www.leica-microsystems.com/4330 Conservation For more than a year from August 2007 through October 2008 museum visitors of the Statens Museum for Kunst, the Danish National Gallery in Copenhagen, were able to experience an open conservation studio in the exhibition area. The reason for bringing the conservators and all their equipment into the exhibition rooms of the museum was the conservation, restoration and technical research of Jacob Jordaens’ (1593 – 1678) early masterpiece “The Tribute Money. Peter Finding the Silver Coin in the Mouth of the Fish”, also known as “The Ferry Boat to Antwerp”. https://www.leica-microsystems.com//science-lab/restoration-in-an-open-workshop/ Mon, 14 Dec 2009 23:00:00 +0000 Anne Haack Christensen, Troels Filtenborg, Dr. Jørgen Wadum https://www.leica-microsystems.com/4332 Conservation Stereo Microscopy On her way to work, Ruth Klebel is often approached by tourists asking for the times of guided tours. She always gives the same answer before disappearing behind the wide automatic gate: “I’m afraid there aren’t any, this is private property.” As a restorer of the collections of the Prince of Liechtenstein, Klebel is one of the very few people who regularly come and go at Vaduz Castle without actually living here. https://www.leica-microsystems.com//science-lab/restoring-former-glory-with-cotton-buds-and-a-microscope-the-princely-collections-of-liechtenstein/ Mon, 14 Dec 2009 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, M.A. Ruth Klebel https://www.leica-microsystems.com/4334 Conservation Digital Microscopy Everyone knows that antique marble sculptures were white. Or were they? Scientists of the Copenhagen Polychromy Network (CPN) help to show that the statues of the Greeks and Romans were decorated with extravagant ornaments and sumptuous colours. With the help of a surgical microscope and digital microscopy, the conservators detect tiny traces of paint pigment that suggest a veritable feast of colour in ancient times. https://www.leica-microsystems.com//science-lab/ancient-feast-of-color/ Mon, 14 Dec 2009 23:00:00 +0000 Jan Stubbe Ostergaard https://www.leica-microsystems.com/4341 Conservation Stereo Microscopy In 1852 Leopoldo Alinari, with his brothers Giuseppe and Romualdo, founded a photographic workshop in Florence, which is at the heart of the firm that still bears his name: Fratelli Alinari. It was the beginning of a unique endeavour that specialised in photographic portraiture, works of art and historical monuments, achieving national and international recognition. https://www.leica-microsystems.com//science-lab/save-our-memory/ Mon, 14 Dec 2009 23:00:00 +0000 Sam Habibi Minelli https://www.leica-microsystems.com/9462 Surgical Microscopy People who can’t hear are outsiders, cut off from normal conversation. Children born into a silent world never learn to talk. Adults who lose their hearing due to age, accident or illness are no longer able to participate in social life as they did before and are moreover often branded as slow-witted. Prof. Dr. med. Jan Maurer, Senior Consultant of the ENT clinic and Medical Director of the Catholic Hospital of Koblenz, Germany, has specialised, among other things, in special hearing implants such as the cochlea implant, and post-implantation rehabilitative therapy. https://www.leica-microsystems.com//science-lab/cochlea-implants-for-the-deaf-and-severely-hard-of-hearing/ Tue, 01 Dec 2009 16:47:00 +0000 Dipl. oec.-troph. Anja Schué, Prof. Jan Maurer https://www.leica-microsystems.com/3833 Laser Microdissection Neuroscience Why do we grow old? Research scientists have been looking for an answer to this question for many years – particularly against the background of the increase in neurodegenerative diseases among older people such as Morbus Parkinson. https://www.leica-microsystems.com//science-lab/the-mitochondrial-hypothesis-of-ageing/ Mon, 07 Sep 2009 16:11:00 +0000 Dr. Andreas Bender, Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/15667 Light Sheet Microscopy Selective plane illumination microscopy (SPIM) and other fluorescence microscopy techniques in which a focused sheet of light serves to illuminate the sample have become increasingly popular in developmental studies. Fluorescence light-sheet microscopy bridges the gap in image quality between fluorescence stereomicroscopy and high-resolution imaging of fixed tissue sections. https://www.leica-microsystems.com//science-lab/selective-plane-illumination-microscopy-techniques-in-developmental-biology/ Mon, 15 Jun 2009 11:27:00 +0000 https://www.leica-microsystems.com/9458 Surgical Microscopy We’ve all experienced an everyday product at some time that had great technology and a stylish design but was totally impractical to use. A designer coffee pot that spills its contents all round the cup or a mobile phone with such tiny keys that you can’t help pressing two at once. If instruments for medical professionals were designed with so little regard for practical application, they’d have no chance of surviving on the market. https://www.leica-microsystems.com//science-lab/just-what-the-surgeon-wants/ Fri, 29 May 2009 11:39:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/9456 Surgical Microscopy Cataract surgery has been practised for centuries and is one of the most common operations performed worldwide today. Ultra modern surgical techniques with tiny incisions and high-quality prosthetic lenses make the operation extremely safe and yield excellent results. Nevertheless, cataract surgery is being perfected all the time. https://www.leica-microsystems.com//science-lab/seeing-without-a-haze-trends-in-cataract-surgery/ Fri, 29 May 2009 10:59:00 +0000 Dipl. oec.-troph. Anja Schué, Dr. Antoine P. Brézin https://www.leica-microsystems.com/2558 Stereo Microscopy Basics in Microscopy Widefield Microscopy Manufacturers digital cameras race to outdo each other with ever-increasing numbers of megapixels. The world record for professional medium format digital cameras has now surpassed 60 megapixels per shot using a very large and expensive sensor with a resolution of about 9000 x 6700 pixels. Each time you capture such an image you get about 180 MB of uncompressed data and even more if you switch to 16-bit per colour for full dynamic range. https://www.leica-microsystems.com//science-lab/digital-cameras/ Thu, 30 Apr 2009 22:00:00 +0000 Urs Schmid https://www.leica-microsystems.com/8076 Surface Metrology Digital Microscopy Quality Assurance Solar energy is becoming more and more important all around the globe. Not only is it available in unlimited supply, it also offers key advantages for protecting the climate and the environment. Every year, many thousands of solar cells are produced worldwide for new photovoltaic plants. An important criterion for quality control is 3D characterization of the light-absorbing surface. In the past, this required time-consuming SEM analysis. A Dual-Core 3D Measuring Microscope that combines confocal and interferometry technology offers non-contact, high-precision analysis of the surface texture of solar cells in a matter of seconds. https://www.leica-microsystems.com//science-lab/3d-surface-measurement-characterization-of-solar-cells/ Mon, 27 Apr 2009 22:00:00 +0000 Roger Artigas https://www.leica-microsystems.com/11047 Metallography Quality Assurance Steel, an alloy of iron and carbon, is both stable and elastic, extremely resistant, and a permanent item in our everyday life. Today there are over 2,500 standard steel types, with new grades and applications emerging all the time. Each steel type is specially made for its purpose. It is subject to stringent quality standards to ensure that it optimally withstands the specific loads. https://www.leica-microsystems.com//science-lab/steel-it-all-depends-on-whats-really-inside/ Wed, 15 Apr 2009 11:34:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/8078 Surface Metrology Quality Assurance Digital Microscopy Micro-optic components are used in a host of products concerned with illumination or imaging. Quality control of these components is challenging, as inspection has to be highly accurate yet non-invasive. Combining confocal and interferometric technology, the Leica DCM 3D Measuring Microscope is equipped to characterize these components with nanometer precision and ultra fast speed, despite the fact that they often have polished and curved surfaces that are difficult to examine. https://www.leica-microsystems.com//science-lab/no-defect-goes-undetected-inspection-of-micro-optic-components/ Tue, 14 Apr 2009 22:00:00 +0000 Roger Artigas https://www.leica-microsystems.com/8082 Quantitative Imaging Widefield Microscopy Förster Resonance Energy Transfer (FRET) is a technique, which allows insight into the interactions between proteins or molecules in proximities beyond light microscopic resolution. An excited fluorophore, called the donor, transfers its excited state energy to a light absorbing molecule which is called the acceptor. This transfer of energy is non-radiative. Sensitized Emission is one established method for the evaluation of FRET efficiencies. It can be applied to live cells as well as to fixed samples. https://www.leica-microsystems.com//science-lab/fret-sensitized-emission-wizard-widefield/ Tue, 14 Apr 2009 22:00:00 +0000 Markus Schechter, Gabriele Burger https://www.leica-microsystems.com/8111 Neuroscience Live-Cell Imaging The function of nerve and muscle cells relies on ionic currents flowing through ion channels. These ion channels play a major role in cell physiology. One way to investigate ion channels is to use patch clamping. This method allows investigation of ion channels in detail and recording of the electric activity of different types of cells, mainly excitable cells like neurons, muscle fibres or beta cells of the pancreas. The patch clamping technique was developed by Erwin Neher and Bert Sakmann in the 1970s and 80s to study individual ion channels in living cells. In 1991 they received the Nobel Prize for Physiology and Medicine for their work. Today the patch clamping technique is one of the most important methods in the field of electrophysiology. https://www.leica-microsystems.com//science-lab/new-standard-in-electrophysiology-and-deep-tissue-imaging/ Tue, 17 Mar 2009 22:04:00 +0000 Dr. Irmtraud Steinmetz https://www.leica-microsystems.com/9445 Super-Resolution We demonstrate beam scanning-stimulated emission depletion microscopy with in vivo labeled cells. A red emitting fluorescent dye is introduced into membrane protein fused to a multifunctional reporter protein (HaloTag, Promega, Madison, WI) in live cells. This approach allows superresolution stimulated emission depletion imaging without the limitations of immunofluorescence-based staining. https://www.leica-microsystems.com//science-lab/in-vivo-labeling-method-using-a-genetic-construct-for-nanoscale-resolution-microscopy/ Wed, 07 Jan 2009 12:21:00 +0000 Dr. Jan Schröder, Hélène Benink, Marcus Dyba https://www.leica-microsystems.com/8080 Surface Metrology Quality Assurance In recent years, interferometers and optical imaging profilers based on confocal technology have been competing fiercely to conquer the non-contact surface metrology market. They are both capable of accurately and reliably measuring surface topographies on a millimeter to nanometer scale. Leica Microsystems presents a complete solution which combines both confocal and interferometry techniques: the Leica DCM 3D Dual-Core 3D Measuring Microscope. https://www.leica-microsystems.com//science-lab/3d-measuring-microscope-combines-confocal-and-interferometry-techniques/ Wed, 19 Nov 2008 23:00:00 +0000 Roger Artigas https://www.leica-microsystems.com/10319 CLEM EM Sample Preparation Correlative light/electron microscopy (CLEM) allows the simultaneous observation of a given subcellular structure by fluorescence light microscopy (FLM) and electron microscopy. The use of this approach is becoming increasingly frequent in cell biology. In this study, we report on a new high data output CLEM method based on the use of cryosections. https://www.leica-microsystems.com//science-lab/high-data-output-and-automated-3d-correlative-light-electron-microscopy-method/ Sat, 15 Nov 2008 12:52:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/5151 Metallography To see how liquids can be made to flow, without being directly heated or touched, you only have to watch a raw egg explode in a microwave oven. Electromagnetic forces can even melt metal at hotter than 1000 °C. In the Magnetohydrodynamics study group at the Research Centre Dresden-Rossendorf (FZD) these complex interactions between electrically conductive liquids and magnetic fields are used to control the flow and solidification processes of liquid metal alloys. https://www.leica-microsystems.com//science-lab/research-for-the-optimal-structure/ Sun, 02 Nov 2008 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, Dr. Sven Eckert https://www.leica-microsystems.com/14848 Confocal Microscopy Epithelial polarity is based on intracellular sorting machinery that maintains the asymmetric distribution of lipids and proteins to the cell surface. Dependent on their lipid raft affinity, newly synthesized apical polypeptides are segregated into distinct vesicle populations subsequent to the passage through the Golgi apparatus. https://www.leica-microsystems.com//science-lab/apical-cargo-traverses-endosomal-compartments-on-the-passage-to-the-cell-surface/ Mon, 15 Sep 2008 17:25:00 +0000 https://www.leica-microsystems.com/14660 Super-Resolution Our interview partner on the topic of Pioneering is Prf. Stefan Hell, a scientific member of the Max Planck Society and director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, where he heads the Department of Nanobiophotonics. In addition to further chairs and memberships, he is also head of the High resolution Optical Microscopy research group, a partnership department of the German Cancer Research Center (DFKZ). https://www.leica-microsystems.com//science-lab/interview-with-stefan-hell-for-me-pioneering-is/ Sun, 31 Aug 2008 22:00:00 +0000 https://www.leica-microsystems.com/9421 Confocal Microscopy Until now, biological and medical research fluorescence imaging in multi-user facilities or institutes has been limited by the type or number of dyes that could be excited. The Leica TCS SP5 X supercontinuum confocal unites the broadband capabilities of the Leica TCS SP5 AOBS® and the freedom and flexibility to select any excitation line within the continuous range of 470 to 670 nm. https://www.leica-microsystems.com//science-lab/the-first-supercontinuum-confocal-that-adapts-to-the-sample/ Fri, 22 Aug 2008 16:40:00 +0000 PhD Kees Jalink, Prof. Alberto Diaspro, Valentina Caorsi, Paolo Bianchini, Dr. Rolf T. Borlinghaus, Scott Young https://www.leica-microsystems.com/4290 Multiphoton Microscopy Confocal Microscopy Quantitative Imaging Cancer research using confocal and multiphoton microscopy. Sequencing of the human genome stimulated a radical change in the approach to biomedical research. The comprehension of the mechanisms regulating life gained a scale-up in throughput to speed up the retrieval of data for a global vision of a system of incomparable complexity. https://www.leica-microsystems.com//science-lab/from-molecules-to-tissues/ Thu, 21 Aug 2008 22:00:00 +0000 Mario Faretta, Davide Mazza, Pietro Transidico, Simona Ronzoni, Dario Parazzoli, Andrea Palamidessi, Ivan Muradore, Massimiliano Garrè, Sara Barozzi, Prof. Alberto Diaspro, Ph.D. Illaria Testa https://www.leica-microsystems.com/4303 Confocal Microscopy Lipids are on everyone’s lips nowadays, whether ω-3/6 fatty acids, good and bad cholesterol or just plain fat that has the annoying habit of accumulating on our hips. Serious diseases such as obesity, arteriosclerosis and type 2 diabetes mellitus are directly connected with lipid metabolism disorders. https://www.leica-microsystems.com//science-lab/obese-and-slim-yeast-cells/ Thu, 21 Aug 2008 22:00:00 +0000 Univ.-Prof., Dipl. Ing., Dr. techn. Sepp Dieter Kohlwein https://www.leica-microsystems.com/9116 Forensics Wayne Buttermore, Manager Regional Sales Industry at Leica Microsystems, interviewed Skip Palenik to hear his perspective on the benefits of continuing education in forensic science. https://www.leica-microsystems.com//science-lab/the-future-of-forensic-training-programs/ Wed, 06 Aug 2008 22:00:00 +0000 Wayne Buttermore, Skip Palenik https://www.leica-microsystems.com/9122 Forensics The impact of modern scientific methods on the analysis of crime scene evidence has dramatically changed many forensic sub-specialties. Arguably one of the most dramatic examples is the impact of molecular biology on the analysis of biological evidence. https://www.leica-microsystems.com//science-lab/forensic-detection-of-sperm-from-sexual-assault-evidence/ Wed, 06 Aug 2008 22:00:00 +0000 Ph.D., CSO Karl Reich https://www.leica-microsystems.com/10190 Quantitative Imaging Multiphoton Microscopy Quantitative measurement method based on FRAP and FRAPa using multiphoton microscopy. We present a new convenient method for quantitative three-dimensionally resolved diffusion measurements based on the photobleaching (FRAP) or photoactivation (FRAPa) of a disk-shaped area by the scanning laser beam of a multiphoton microscope. Contrary to previously reported spot-photobleaching protocols, this method has the advantage of full scalability of the size of the photobleached area and thus the range of diffusion coefficients, which can be measured conveniently. https://www.leica-microsystems.com//science-lab/a-new-frapfrapa-method-for-three-dimensional-diffusion-measurements-based-on-multiphoton-excitation-microscopy/ Fri, 11 Jul 2008 15:06:00 +0000 Davide Mazza https://www.leica-microsystems.com/2472 Basics in Microscopy Widefield Microscopy This article explains how to avoid the phenomen of "empty magnification" in microscopy. https://www.leica-microsystems.com//science-lab/beware-of-empty-magnification/ Sun, 18 May 2008 10:00:00 +0000 Dipl. oec.-troph. Anja Schué https://www.leica-microsystems.com/5340 Quality Assurance An exquisite beverage deserves a high-quality glass. Even the ancient Romans made artistically crafted drinking glasses. In the Middle Ages, Venetian glassmakers were famous for the purity of their glass. One of the oldest materials known to man, glass is used today in many areas, in which it must meet the most stringent quality standards. https://www.leica-microsystems.com//science-lab/quality-as-clear-as-glass-polarizing-microscopy-in-glass-production/ Thu, 17 Apr 2008 22:00:00 +0000 Klaus-Peter Martinek https://www.leica-microsystems.com/5350 Stereo Microscopy Quality Assurance A study carried out jointly by Leica Microsystems and the Institute of Neuroinformatics at the University of Zurich and Swiss Federal Institute of Technology provided the basis for an innovation in stereomicroscopy: FusionOptics™. The significant performance increase attained by FusionOptics™ is highly valuable for everyday work at the microscope. https://www.leica-microsystems.com//science-lab/fusionoptics-combines-high-resolution-and-depth-of-field-for-ideal-3d-optical-images/ Thu, 17 Apr 2008 22:00:00 +0000 Daniel Goeggel, Dipl. oec.-troph. Anja Schué, Dr. Daniel Kiper https://www.leica-microsystems.com/10325 Quantitative Imaging We recently reported on CFP-Epac-YFP, an Epac-based single polypeptide FRET reporter to resolve cAMP levels in living cells. In this study, we compared and optimized the fluorescent protein donor/acceptor pairs for use in biosensors such as CFP-Epac-YFP. Our strategy was to prepare a wide range of constructs consisting of different donor and acceptor fluorescent proteins separated by a short linker. https://www.leica-microsystems.com//science-lab/a-comparison-of-donor-acceptor-pairs-for-genetically-encoded-fret-sensors-application-to-the-epac-camp-sensor-as-an-example/ Wed, 02 Apr 2008 16:09:00 +0000 https://www.leica-microsystems.com/9128 Forensics Wayne Buttermore, Marketing Manager for Forensic Microscopy, Leica Microsystems, had the distinct honor of interviewing Kevin Lothridge, Executive Director of the National Forensic Science Technology Center (NFSTC) to hear his perspectives on continuing education in the forensic science community. https://www.leica-microsystems.com//science-lab/science-serving-justice/ Tue, 01 Apr 2008 22:00:00 +0000 Wayne Buttermore https://www.leica-microsystems.com/10689 Ergonomics This study evaluated the effect of an occupational therapy ergonomics intervention on the workstation design and body positioning of microscope workers at a fiber optics facility. https://www.leica-microsystems.com//science-lab/ergonomics/implementing-ergonomic-design-for-microscope-workstations/ Fri, 01 Feb 2008 15:12:00 +0000 https://www.leica-microsystems.com/3006 Live-Cell Imaging TIRF Microscopy Using TIRF microscopy, scientists have been able to take a closer look at intracellular transport processes with the example of the galactose-binding protein Galectin-3, which has been identified as a potential apical sorting receptor. https://www.leica-microsystems.com//science-lab/exploring-cell-logistics/ Fri, 02 Nov 2007 11:29:00 +0000 Prof. Ralf Jacob, Dipl. Biol. Dominik Schneider https://www.leica-microsystems.com/3397 Stereo Microscopy Confocal Microscopy For the study of minute marine organisms such as free swimming larvae or bottom dwelling invertebrates it is a great advantage to have modern microscopic equipment in close proximity to the natural habitat. This allows the acquisition of images from organisms under virtually natural conditions. https://www.leica-microsystems.com//science-lab/in-the-footsteps-of-linnaeus-microscopic-imaging-in-marine-biodiversity-research/ Thu, 01 Nov 2007 23:00:00 +0000 Ph.D. Sam Dupont, Ph.D. Matthias Obst, Mike Thorndyke https://www.leica-microsystems.com/4071 Confocal Microscopy Artificial illumination can harm hypogean monuments by inducing the uncontrolled growth of photosynthetic biofilms (green sickness or maladie verte). With the aim of preventing biodeterioration or aesthetic damage in Roman hypogean monuments (the St. Callistus and St. Domitilla catacombs, Rome, Italy), a confocal technique is used for the analysis of fluorescent pigments from a single cell, based on spectrofluorometric methods. https://www.leica-microsystems.com//science-lab/characterization-of-photosynthetic-biofilms-from-roman-catacombs-via-3d-imaging-and-subcellular-identification-of-pigments/ Thu, 01 Nov 2007 23:00:00 +0000 Monica Roldán, M. Hernández-Mariné https://www.leica-microsystems.com/9447 Stereo Microscopy Embryonic stem (ES) cells have been investigated as a potential replacement therapy for failed organs, such as the liver. However, detection of hepatic engraftment from candidate stem cells has been difficult due to low engraftment efficiency. Previous detection methods required that the graft be processed by molecular and/or immunohistochemical techniques, limiting further functional studies. This study evaluated the use of three-dimensional fluorescent stereomicroscopy for gross detection of ES cell derived hepatic engraftment. https://www.leica-microsystems.com//science-lab/detection-and-characterization-of-hepatic-engraftment-of-embryonic-stem-derived-cells-by-fluorescent-stereomicroscopy/ Wed, 15 Aug 2007 13:23:00 +0000 https://www.leica-microsystems.com/7970 CLEM EM Sample Preparation In recent years light microscopy studies have been dominated by live cell imaging while electron microscopy has been used for high-resolution studies. Latterly, there has been increasing interest in combining these techniques. This combination is called Correlative Light Electron Microscopy (CLEM). Due to the high resolution made possible by electron microscopy, artefacts induced during preparation of a sample can, however, also be clearly seen. https://www.leica-microsystems.com//science-lab/clem-combining-the-strengths-of-light-and-electron-microscopy/ Mon, 09 Apr 2007 22:00:00 +0000 Dr. Paul Verkade https://www.leica-microsystems.com/9352 Quantitative Imaging Confocal Microscopy In the last decade, fluorescence resonance energy transfer (FRET) has become a useful technique for studying intermolecular interactions applied to the analysis of biological systems. Although FRET measurements may be very helpful in the comprehension of different cellular processes, it can be difficult to obtain quantitative results, hence the necessity of studying FRET on controllable systems. https://www.leica-microsystems.com//science-lab/fret-measurements-on-fuzzy-fluorescent-nanostructures/ Tue, 27 Mar 2007 22:00:00 +0000 Valentina Caorsi, Emiliano Ronzitti, Ph.D. Giuseppe Vicidomini, Prof. Gail McConnell, Prof. Alberto Diaspro https://www.leica-microsystems.com/5139 Stereo Microscopy Until the middle of the 19th century, microscopes were hand-crafted custom instruments. At the time, it was not possible to predict the properties of a lens in advance, so lenses were shaped and tested repeatedly until they delivered the desired magnification. https://www.leica-microsystems.com//science-lab/the-history-of-stereo-microscopy-part-iii/ Thu, 11 Jan 2007 13:03:00 +0000 Daniel Goeggel https://www.leica-microsystems.com/5137 Stereo Microscopy For millennia, glass was admired for its beauty and the artful objects that could be created with it. With the rise of the exact sciences, however, researchers became more demanding with regard to glass quality. Traditional glass was not suitable for microscope lenses – streaks, bubbles and inclusions containing impurities made precise observation impossible. https://www.leica-microsystems.com//science-lab/the-history-of-stereo-microscopy-part-ii/ Thu, 11 Jan 2007 10:18:00 +0000 Daniel Goeggel https://www.leica-microsystems.com/5129 Stereo Microscopy The 17th century in Europe was marked by the Thirty Years' War (1618–1648). The population lived in constant fear and poverty. Marauding hordes wandered the countryside; failed harvests, plagues, pestilence and starvation were the order of the day. Most of the population had little in the way of clothing or shoes, and lived in modest huts infested by parasites and rats. https://www.leica-microsystems.com//science-lab/the-history-of-stereo-microscopy-part-i/ Wed, 10 Jan 2007 23:00:00 +0000 Daniel Goeggel https://www.leica-microsystems.com/4394 Confocal Microscopy Quantitative Imaging Multiphoton Microscopy We report about two-photon activation of a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (pa-GFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm. Two-photon photoactivation produces an effective real three-dimensional (3D) localization of the molecular switching of pa-GFP in the bright state. https://www.leica-microsystems.com//science-lab/4d-photoactivation-of-pa-gfp-in-living-cells-using-two-photon-excitation-laser-scanning-microscopy/ Sat, 14 Oct 2006 22:00:00 +0000 Prof. Alberto Diaspro, Massimiliano Garrè, Pietro Transidico, Dario Parazzoli, Sara Barozzi, Ph.D. Giuseppe Vicidomini, Davide Mazza, Valentina Caorsi, Ph.D. Illaria Testa, Mario Faretta https://www.leica-microsystems.com/8084 Quantitative Imaging Confocal Microscopy Förster Resonance Energy Transfer (FRET) is a technique, which allows insight into the interactions between proteins or molecules in proximities beyond light microscopic resolution. https://www.leica-microsystems.com//science-lab/fret-sensitized-emission-wizard-confocal/ Thu, 14 Sep 2006 22:00:00 +0000 Markus Schechter, Gabriele Burger https://www.leica-microsystems.com/6771 Multiphoton Microscopy Confocal Microscopy Advanced microscopical techique for life science: multiphoton microscopy. Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. https://www.leica-microsystems.com//science-lab/multi-photon-excitation-microscopy/ Mon, 05 Jun 2006 22:00:00 +0000 Prof. Alberto Diaspro, Paolo Bianchini, Ph.D. Giuseppe Vicidomini, Mario Faretta, Paola Ramoino, Cesare Usai https://www.leica-microsystems.com/10023 Quantitative Imaging Among all photobleaching experiments which have been described, fluorescence recovery after photobleaching (FRAP) is the most popular. It employs irradiation of a fluorophore in a living sample with a short laser pulse to degrade it and thereby abolish fluorescence followed by time-resolved image recording of the sample. https://www.leica-microsystems.com//science-lab/fluorescence-recovery-after-photobleaching-with-the-leica-tcs-sp2/ Sun, 15 Aug 2004 19:15:00 +0000 Dr. Constantin Kappel, Professor Dr. Roland Eils https://www.leica-microsystems.com/10327 Quantitative Imaging Confocal Microscopy Imaging of fluorescence resonance energy transfer (FRET) between suitable fluorophores is increasingly being used to study cellular processes with high spatiotemporal resolution. The genetically encoded Cyan (CFP) and Yellow (YFP) variants of Green Fluorescent Protein have become the most popular donor and acceptor pair in cell biology. FRET between these fluorophores can be imaged by detecting sensitized emission. This technique, for which CFP is excited and transfer is detected as emission of YFP, is sensitive, fast, and straightforward, provided that proper corrections are made. In this study, the detection of sensitized emission between CFP and YFP by confocal microscopy is optimized. https://www.leica-microsystems.com//science-lab/correcting-confocal-acquisition-to-optimize-imaging-of-fluorescence-resonance-energy-transfer-by-sensitized-emission/ Tue, 20 Apr 2004 17:02:00 +0000 PhD Jacco van Rheenen https://www.leica-microsystems.com/9682 Fluorescence Microscopy Fluorescence is a process where a substance after having absorbed light (photons) emitts a radiation the wavelength (colour) of which is longer than that of the absorbed light, and where this emission stops immediately after cessation of the excitation. This phenomenon is the basic element of fluorescence microscopy and its application. https://www.leica-microsystems.com//science-lab/multi-wavelength-epi-illumination-in-fluorescence-microscopy/ Sat, 01 Dec 2001 15:35:00 +0000 Professor em. Johan Sebastiaan Ploem, Dr. Friedrich Walter