Leica Science Lab - Tag : Confocal Microscopy https://www.leica-microsystems.com//science-lab/tag/tags/confocal-microscopy/show/Tag/ Article tagged with Confocal Microscopy en-US https://www.leica-microsystems.com/29648 Confocal Microscopy Explore Innovative Techniques to Separate Fluorophores with Overlapping Spectra 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/29614 Confocal Microscopy Add an Extra Dimension to Confocal Results with TauSense 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/28460 Confocal Microscopy How to Uncover Hidden Dimensions in Research with Lifetime Imaging 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/28307 Confocal Microscopy Learn how to Remove Autofluorescence from your Confocal Images 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/26903 Confocal Microscopy Pore Scale Visualization of Drainage in 3D Porous Media by 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/26677 Quantitative Imaging SP8 FALCON: a novel concept in fluorescence lifetime imaging enabling video-rate confocal FLIM 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/4421 Coherent Raman Scattering (CRS) Coherent Raman Scattering Microscopy Publication List 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/26318 Neuroscience Confocal Microscopy Cellular Motility: Microtubules, Motor Proteins and Tau-Proteins 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/25023 Confocal Microscopy Visualize developmental patterns in Drosophila 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/24929 Confocal Microscopy Visualize mechanical interactions of Cancer Cells 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 Zebrafish Brain - Whole Organ Imaging at High Resolution 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/24698 Confocal Microscopy What is a Resonant Scanner? 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/24696 Confocal Microscopy What is a Spectral Detector (SP Detector)? 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/24709 Confocal Microscopy What is a Field-of-View Scanner? 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/24694 Confocal Microscopy What is a Tandem Scanner? 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/24562 Confocal Microscopy Kaggle Competition for Multi-label Classification of Cell Organelles in Proteome Scale Human Protein Atlas Data 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/24560 Super-Resolution See More Than Just Your Image 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 Dynamic Endoplasmic Reticulum 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/20671 Confocal Microscopy MONENSIN SENSITIVITY1 (MON1)/CALCIUM CAFFEINE ZINC SENSITIVITY1 (CCZ1) 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/20323 Confocal Microscopy Which Sensor is the Best for Confocal Imaging? 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/20297 Confocal Microscopy Image Gallery: FLIM for high-speed investigation of molecular interactions 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/20175 Confocal Microscopy Widefield Microscopy mTORC1 Promotes Proliferation of Immature Schwann Cells and Myelin Growth of Differentiated Schwann Cells 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/20133 Quantitative Imaging FCS - Fluorescence Correlation Spectroscopy 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 FLIM FRET - Förster Resonance Energy Transfer 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 FLCS - Fluorescence Lifetime Correlation Spectroscopy 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 What is FLIM - Fluorescence Lifetime 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 SP FLIM - Spectral Fluorescence Lifetime 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 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/19990 Confocal Microscopy IT Security Whitepaper 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 Expression Analysis of Platelet‐derived Growth Factor Receptor Alpha and its Ligands in the Developing Mouse Lung 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/19872 Super-Resolution Abstracts of the 7th European Super-Resolution User-Club Meeting 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/19283 Confocal Microscopy Primary Beam Splitting Devices for Confocal Microscopes 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/19321 Confocal Microscopy Pinhole Effect in Confocal Microscopes 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/19111 Confocal Microscopy Light Sheet Microscopy Multiphoton Microscopy Neuroscience Clarifying Tissue Clearing 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/18994 Fluorescence Microscopy Live-Cell Imaging Widefield Microscopy Chronic Inflammation Under the Microscope 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/18925 Super-Resolution Confocal Microscopy Methods to Calibrate and Scale Axial Distances in Confocal Microscopy as a Function of Refractive Index 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/18935 Fluorescence Microscopy Confocal Microscopy Multispectral Phloem-Mobile Probes: Properties and Applications 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/18900 Super-Resolution Confocal Microscopy P53- and Mevalonate Pathway–Driven Malignancies Require Arf6 for Metastasis and Drug Resistance 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/18730 Confocal Microscopy Live-Cell Imaging Adeno-associated Viral Vectors do not Efficiently Target Muscle Satellite Cells 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/17673 Confocal Microscopy Super-Resolution HyVolution – Super-Resolution Imaging with a Confocal Microscope 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/17669 Confocal Microscopy Super-Resolution HyVolution – the Smart Path to Confocal 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/13262 Quantitative Imaging FRAP with TCS SP8 Resonant Scanner 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/16587 Light Sheet Microscopy Live-Cell Imaging Confocal Microscopy Light Sheet Microscopy Turned Vertically 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/15426 Confocal Microscopy Progressive Glucose Stimulation of Islet Beta Cells Reveals a Transition From Segregated to Integrated Modular Functional Connectivity Patterns 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/16460 Confocal Microscopy Multiphoton Microscopy From Light to Mind: Sensors and Measuring Techniques in Confocal 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/16099 Confocal Microscopy "We can go home and the imaging is done automatically by the Leica HCS A Matrix Screener." 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/14978 Super-Resolution Confocal Microscopy Detailed Morphological Characterisation of Hendra Virus Infection of Different Cell Types Using Super-Resolution and Conventional Imaging 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 Confocal and Digital Light Sheet Imaging 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/14702 Education Confocal Microscopy Venturing into Uncharted Dimensions – the Fascination of Future Technologies 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/15203 Confocal Microscopy Live-Cell Imaging Video: High Speed Scanning – With two Scanners in one System 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/14884 Confocal Microscopy Super-Resolution Live-Cell Imaging Multiphoton Microscopy Fluorescence Microscopy Quantitative Imaging Neuroscience How to Choose the Right Confocal Microscope for Your Lab? 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/14063 Confocal Microscopy Multiphoton Microscopy Smart Control for Resonant Galvo Scanners 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/13490 Surface Metrology Characterization of Thin Films Using High Definition Confocal Microscopy 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/12420 Confocal Microscopy Live-Cell Imaging Functional Connectivity in Islets of Langerhans from Mouse Pancreas Tissue Slices 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/12648 Surface Metrology Quality Assurance Digital Microscopy 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 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/12002 Confocal Microscopy Live-Cell Imaging The Relationship Between Membrane Potential and Calcium Dynamics in Glucose-Stimulated Beta Cell Syncytium in Acute Mouse Pancreas Tissue Slices 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/11235 Confocal Microscopy Acousto Optics in True Confocal Spectral Microscope Systems 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/9680 Confocal Microscopy Pinhole Geometry: Four Corners are Perfect 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 Step by Step Guide for FRAP Experiments 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 Automated Digital 3D Topography Measurement with the Leica DCM 3D Dual Core Measuring Microscope 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/7246 Multiphoton Microscopy Developments in Multiphoton Excitation 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/5082 Confocal Microscopy Quantitative Imaging FLCS – Advances in Fluorescence Correlation Spectroscopy 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/4487 Confocal Microscopy Beam Splitting 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 Spectral Imaging 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/4718 Neuroscience Neurobiology and Microscopy 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 Förster Resonance Energy Transfer (FRET) 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) An Introduction to CARS Microscopy 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/4384 Confocal Microscopy Mosaic Images 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/4126 Confocal Microscopy Fluorescence Microscopy Brighter Fluorescence by Resonant Scanning 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/3772 Confocal Microscopy Pinhole Controls Optical Slicing 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 Optical Section Thickness 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 Lasers for Confocal 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/2860 Confocal Microscopy Confocal Microscopy - Optical Path “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/3514 Confocal Microscopy Quantitative Imaging Choose Your Excitation Wavelength 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/2471 Neuroscience Super-Resolution The Missing Link to the Nanocosm of Life 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 Restless Receptors 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/2731 Confocal Microscopy Multiphoton Microscopy What is an OPO? 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/2743 Coherent Raman Scattering (CRS) Multiphoton Microscopy Confocal Microscopy CARS and Confocal 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/2730 Multiphoton Microscopy Confocal Microscopy Deep Tissue Imaging 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/2732 Confocal Microscopy Multiphoton Microscopy Live-Cell Imaging Neuroscience Exploring the Concert of Neuronal Activities 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/2561 Multiphoton Microscopy Live-Cell Imaging Confocal Microscopy Stem Cell Biology in Cancer Research 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 Confocal Nanoscopy Goes Multicolor 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/8080 Surface Metrology Quality Assurance 3D Measuring Microscope Combines Confocal and Interferometry Techniques 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/4303 Confocal Microscopy Obese and Slim Yeast Cells 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/4290 Multiphoton Microscopy Confocal Microscopy Quantitative Imaging From Molecules to Tissues 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/4071 Confocal Microscopy Characterization of Photosynthetic Biofilms from Roman Catacombs via 3D Imaging and Subcellular Identification of Pigments 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/3397 Stereo Microscopy Confocal Microscopy In the Footsteps of Linnaeus – Microscopic Imaging in Marine Biodiversity Research 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/4394 Confocal Microscopy Quantitative Imaging Multiphoton Microscopy 4D Photoactivation of pa-GFP in Living Cells Using Two-Photon Excitation Laser Scanning 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 FRET Sensitized Emission Wizard Confocal 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 Multi-photon Excitation 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 Fluorescence Recovery after Photobleaching with the Leica TCS SP2 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