Leica Science Lab - Tag : Live Imaging https://www.leica-microsystems.com//science-lab/tag/live-imaging/ Article tagged with Live Imaging en-US https://www.leica-microsystems.com/72888 Light Sheet Microscopy Following Ca²⁺ Changes in Acutely Stimulated Spheroids Spheroids are easy and cost-effective 3D cell cultures, useful for fundamental and applied research. They can bridge the gap between 2D and in vivo experiments and can be used for high-throughput screenings. However, performing live microscopy to visualize acute drug responses is difficult with these roundish and non-adherent cell aggregates, hampering the study of rapid changes in intra- and intercellular signalling. To solve this issue, we stabilized spheroids in micro-perfusion chambers using a transparent and soft gelatine mesh. https://www.leica-microsystems.com/science-lab/following-ca2-changes-in-acutely-stimulated-spheroids/ Wed, 23 Jun 2021 12:16:00 +0000 https://www.leica-microsystems.com/48615 THUNDER Imaging Fluorescence Microscopy Widefield Microscopy Improvement of Imaging Techniques to Understand Organelle Membrane Cell Dynamics Understanding cell functions in normal and tumorous tissue is a key factor in advancing research of potential treatment strategies and understanding why some treatments might fail. Single-cell analysis is crucial in biomedical research to unravel which cellular and molecular pathways are altered in complex diseases such as cancer. https://www.leica-microsystems.com/improvement-of-imaging-techniques-to-understand-organelle-membrane-cell-dynamics/ Mon, 15 Feb 2021 12:11:00 +0000 Corporate Communications https://www.leica-microsystems.com/27674 Confocal Microscopy Coherent Raman Scattering (CRS) How to prepare samples for Stimulated Raman Scattering 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/4592 Coherent Raman Scattering (CRS) Step by Step Guide to the Molecular Basics of CARS Microscopy 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/27386 Super-Resolution New Light Shed on the Nanodomain Organization of the Endoplasmic Reticulum (ER) 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/26832 Quantitative Imaging Imaging Intracellular Temperature using Fluorescence Lifetime Imaging Microscopy (FLIM) 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/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/26294 Quantitative Imaging Development of fluorescence lifetime imaging microscopy (FLIM) and its relevance for functional 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/24669 Super-Resolution Extending Nanoscopy Possibilities with STED and exchangeable fluorophores 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/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/20653 Laser Microdissection Live Cell Isolation by 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/20061 Live-Cell Imaging Solvent immersion imprint lithography 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/19922 Fluorescence Microscopy Widefield Microscopy Acute Transcriptional Up-regulation Specific to Osteoblasts/Osteoclasts in Medaka Fish Immediately after Exposure to Microgravity 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/19686 Live-Cell Imaging Introduction to Mammalian Cell Culture 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/19342 Super-Resolution Axial Tubule Junctions Control Rapid Calcium Signaling in Atria 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/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/18278 Super-Resolution Live-Cell Imaging Super-Resolution Optical Microscopy of Lipid Plasma Membrane Dynamics 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/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/15428 Confocal Microscopy Live-Cell Imaging Highly Selective Fluorescent and Colorimetric Probe for Live-cell Monitoring of Sulphide Based on Bioorthogonal Reaction 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/15645 Confocal Microscopy Live-Cell Imaging Label-free in vivo Imaging of Myelinated Axons in Health and Disease with Spectral Confocal Reflectance Microscopy 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/18431 Quantitative Imaging Live-Cell Imaging Individual Macromolecule Motion in a Crowded Living Cell 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/17977 Confocal Microscopy Live-Cell Imaging The Bimodally Expressed MicroRNA miR‐142 Gates Exit from Pluripotency 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/17931 Live-Cell Imaging What Makes sCMOS Microscope Cameras so Popular? 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/17667 Live-Cell Imaging Widefield Microscopy How to do a Proper Cell Culture Quick Check 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/17611 Super-Resolution Multiphoton Microscopy Live-Cell Imaging 4Pi-RESOLFT Nanoscopy 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/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 Confocal Microscopy Light Sheet Microscopy Live-Cell Imaging 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/16521 Super-Resolution Live-Cell Imaging Fluorescence Microscopy Probes that FIT RNA 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/16174 Super-Resolution Quantitative Imaging Live-Cell Imaging Cortical Actin Networks Induce Spatio-temporal Confinement of Phospholipids in the Plasma Membrane – A Minimally Invasive Investigation by STED-FCS 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/15241 Super-Resolution Live-Cell Imaging Multi-protein Assemblies Underlie the Mesoscale Organization of the Plasma Membrane 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/13642 CLEM EM Sample Preparation Correlative Light and Electron Microscopy – Get Your Free CLEM e-Book for Download 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 Professor Paul Verkade https://www.leica-microsystems.com/15430 Confocal Microscopy Multiphoton Microscopy Live-Cell Imaging Third Harmonic Generation Microscopy – Label-Free 3D-Tissue Imaging and Blood Flow Characterization 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/15111 Super-Resolution Universal PAINT – Dynamic Super-Resolution Microscopy 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/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/15077 Light Sheet Microscopy Nature Methods: Light-sheet Fluorescence Microscopy - Method of the Year 2014 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/15219 Confocal Microscopy "Leica is always flexible and dynamic" - Interview with Audrey Salles, Pasteur Institute, Paris 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/14367 Confocal Microscopy Neuroscience Live-Cell Imaging Fluorescence Microscopy Axon Injury and Regeneration in the Adult Drosophila 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/13911 Super-Resolution Live-Cell Imaging Nanoscale Protein Diffusion by STED-Based Pair Correlation Analysis 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/13950 Super-Resolution Neuroscience STED Microscopy of Living Cells – New Frontiers in Membrane and Neurobiology 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/14066 Multiphoton Microscopy Live-Cell Imaging The Environment Makes the Stem Cell 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/11133 Laser Microdissection Laser Microdissection (LMD) and Fancy Applications 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 Confocal Microscopy Live-Cell Imaging Super-Resolution Neuroscience Fluorescence Microscopy Live-Cell Imaging Evolves to Find New Niches 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/11038 Confocal Microscopy Live-Cell Imaging Nobel Prize 2013 in Physiology or Medicine for Discoveries of the Machinery Regulating Vesicle Traffic 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/10924 Confocal Microscopy Live-Cell Imaging A Flip of the Mitotic Spindle has Disastrous Consequences for Epithelial Cells 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/10447 Confocal Microscopy Laser Microdissection Live-Cell Imaging EM Sample Preparation From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron 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/10653 Super-Resolution Live-Cell Imaging Watching Molecule Movements in Live Cells 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/10290 Super-Resolution Live-Cell Imaging Neuroscience New Labeling Tools Can Help to Realize the Full Potential of Super-Resolution Microscopy 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/10281 Confocal Microscopy CLEM EM Sample Preparation Live-Cell Imaging A Precise and Rapid Mapping Protocol for Correlative Light and Electron Microscopy of small invertebrate organisms 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 Direct In Vivo Evidence for Tumor Propagation by Glioblastoma Cancer Stem Cells 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/9492 Live-Cell Imaging Fluorescence Microscopy Quantitative Imaging Imaging Enzymes at Work 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/15675 Light Sheet Microscopy Light Sheet Fluorescence Microscopy: Beyond the Flatlands 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/6499 Fluorescence Microscopy Live-Cell Imaging Widefield Microscopy Digital Camera Technologies for Scientific Bio-Imaging 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/5945 TIRF Microscopy Controlling the TIRF Penetration Depth is Mandatory for Reproducible Results 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/5106 Live-Cell Imaging Introduction to 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/5108 Confocal Microscopy Live-Cell Imaging Live-cell Imaging Techniques 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/5217 Confocal Microscopy Fluorescence Microscopy Live-Cell Imaging Neuroscience TIRF Microscopy Quantitative Imaging The New Repository on the Block 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/4758 Live-Cell Imaging Neuroscience Ratiometric Imaging 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/10159 Super-Resolution Two-color STED Microscopy of Living Synapses using a Single Laser-beam Pair 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 Quantitative Imaging in Cell Biology: 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/4727 Neuroscience Live-Cell Imaging The Patch-Clamp Technique 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/10165 Super-Resolution Neuroscience Live-Cell Imaging STED Nanoscopy of Actin Dynamics in Synapses deep inside Living Brain Slices 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/10218 Super-Resolution Live-Cell Imaging Nanoscopy in a Living Multicellular Organism Expressing GFP 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 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/2994 Live-Cell Imaging TIRF Microscopy TIRF Microscopy of the Apical Membrane of Polarized Epithelial Cells 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/15669 Light Sheet Microscopy Light Sheet Fluorescence Microscopy - A Review 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/2470 Live-Cell Imaging Neuroscience Sniffing Out the Secrets of Social Behavior 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/15667 Light Sheet Microscopy Selective Plane Illumination Microscopy Techniques in Developmental Biology 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/8082 Quantitative Imaging Widefield Microscopy FRET Sensitized Emission Wizard Widefield 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 New Standard in Electrophysiology and Deep Tissue 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 In Vivo Labeling Method Using a Genetic Construct for Nanoscale Resolution Microscopy 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/3006 Live-Cell Imaging TIRF Microscopy Exploring Cell Logistics 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/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