Leica Science Lab - Tag : Cell Biology https://www.leica-microsystems.com//science-lab/tag/tags/cell-biology/show/Tag/ Article tagged with Cell Biology en-US https://www.leica-microsystems.com/28596 Coherent Raman Scattering (CRS) Vibrational Spectroscopic Imaging in Cells, Tissues and Model Organisms Here we present a webinar on the emerging applications of Stimulated Raman Scattering Microscopy, covering the following topics: Quantitative spectroscopic imaging of label-free or chemically tagged specimens: From cell and tissue biology to medical research, materials science, and microfabrication; Probing 3D cellular model systems (organoids) and tissues for preclinical & translational research in cancer, neurodegenerative diseases, immunology, and digestive disorders; Multi-modal confocal nonlinear optical microscopy, combining CARS, SRS, Second-Harmonic Generation, and 2-Photon Fluorescence for advanced tissue-based diagnostics; Monitoring tissue penetration of pharmacological and cosmetic compounds https://www.leica-microsystems.com//science-lab/vibrational-spectroscopic-imaging-in-cells-tissues-and-model-organisms/ Wed, 29 Apr 2020 08:46:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/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/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/24680 Live-Cell Imaging Confluency Check with PAULA Cell Imager Many cell-based experiments require cells in a certain state. This can include their morphology,  fluorescent protein expression, and the confluency of the cell layer. Researchers check their cells regularly to determine these attributes. Confluency estimation can be very difficult! This guess work is biased by every individual researcher, and can result in lab error.  https://www.leica-microsystems.com//science-lab/confluency-check-with-paula-cell-imager/ Wed, 20 Feb 2019 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/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/20689 Multiphoton Microscopy Image Gallery: Tunable Multicolor Deep In Vivo Imaging Today’s life science research focusses on complex biological processes, such as the causes of cancer and other human diseases. A deep look into tissues and living specimens is vital to understanding the conditions and mechanisms in cells and finding answers to crucial questions challenging the life sciences. https://www.leica-microsystems.com//science-lab/galleries/image-gallery-tunable-multicolor-deep-in-vivo-imaging/ Thu, 01 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/20478 Super-Resolution California NanoSystems Institute at UCLA Publications A list of the published scientific articles which include work done in the ALMS/MSI Facilities. https://www.leica-microsystems.com//science-lab/cnsi-publication-list/ Sun, 14 Jan 2018 23:00:00 +0000 https://www.leica-microsystems.com/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/19451 Live-Cell Imaging Fluorescence Microscopy Multiphoton Microscopy Five Questions Asked: Prof. Dr. Jacco van Rheenen speaks about the most important considerations when imaging deep into mouse tissue When operating a confocal microscope, or when discussing features and parameters of such a device, we inescapably mention the pinhole and its diameter. This short introductory document is meant to explain the significance of the pinhole for those, who did not want to spend too much time to dig into theory and details of confocal microscopy but wanted to have an idea about the effect of the pinhole. https://www.leica-microsystems.com//science-lab/five-questions-asked-prof-dr-jacco-van-rheenen-speaks-about-the-most-important-considerations-when-imaging-deep-into-mouse-tissue/ Fri, 19 May 2017 05:47:00 +0000 PhD Jacco van Rheenen, Dr. Bernd Sägmüller https://www.leica-microsystems.com/19109 Confocal Microscopy Human NK Cell Development Requires CD56-mediated Motility and Formation of the Developmental Synapse While distinct stages of natural killer (NK) cell development have been defined, the molecular interactions that shape human NK cell maturation are poorly understood. Here we define intercellular interactions between developing NK cells and stromal cells which, through contact-dependent mechanisms, promote the generation of mature, functional human NK cells from CD34+ precursors. We show that developing NK cells undergo unique, developmental stage-specific sustained and transient interactions with developmentally supportive stromal cells, and that the relative motility of NK cells increases as they move through development in vitro and ex vivo. https://www.leica-microsystems.com//science-lab/human-nk-cell-development-requires-cd56-mediated-motility-and-formation-of-the-developmental-synapse/ Wed, 15 Feb 2017 07:36:00 +0000 Ph.D. Emily M. Mace https://www.leica-microsystems.com/18098 Super-Resolution Actin-Dependent Vacuolar Occupancy of the Cell Determines Auxin-Induced Growth Repression The cytoskeleton is an early attribute of cellular life, and its main components are composed of conserved proteins. The actin cytoskeleton has a direct impact on the control of cell size in animal cells, but its mechanistic contribution to cellular growth in plants remains largely elusive. Here, we reveal a role of actin in regulating cell size in plants. The actin cytoskeleton shows proximity to vacuoles, and the phytohormone auxin not only controls the organization of actin filaments but also impacts vacuolar morphogenesis in an actin-dependent manner. https://www.leica-microsystems.com//science-lab/actin-dependent-vacuolar-occupancy-of-the-cell-determines-auxin-induced-growth-repression/ Fri, 03 Feb 2017 08:14:00 +0000 https://www.leica-microsystems.com/19182 EM Sample Preparation Variety of RNAs in Peripheral Blood Cells, Plasma, and Plasma Fractions Human peripheral blood contains RNA in cells and in extracellular membrane vesicles, microvesicles and exosomes, as well as in cell-free ribonucleoproteins. Circulating mRNAs and noncoding RNAs, being internalized, possess the ability to modulate vital processes in recipient cells. In this study, with SOLiD sequencing technology, we performed identification, classification, and quantification of RNAs from blood fractions: cells, plasma, plasma vesicles pelleted at 16,000 https://www.leica-microsystems.com//science-lab/variety-of-rnas-in-peripheral-blood-cells-plasma-and-plasma-fractions/ Mon, 30 Jan 2017 10:22:00 +0000 https://www.leica-microsystems.com/18958 Super-Resolution The Bif-1-Dynamin 2 Membrane Fission Machinery Regulates Atg9-Containing Vesicle Generation at the Rab11-Positive Reservoirs Application example of HyVolution Super-Resolution - Atg9 is a multispanning transmembrane protein that is required for autophagosome formation. During autophagy, vesicles containing Atg9 are generated through an unknown mechanism and delivered to the autophagosome formation sites. We have previously reported that Atg9-containing membranes undergo continuous tubulation and fission during nutrient starvation in a manner dependent on the curvature-inducing protein Bif-1/Sh3glb1. Here, we identify Dynamin 2 (DNM2) as a Bif-1-interacting protein that mediates the fission of Atg9-containing membranes during autophagy. https://www.leica-microsystems.com//science-lab/the-bif-1-dynamin-2-membrane-fission-machinery-regulates-atg9-containing-vesicle-generation-at-the-rab11-positive-reservoirs/ Wed, 25 Jan 2017 13:00:00 +0000 https://www.leica-microsystems.com/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/18806 Super-Resolution Practical Guide for Excellent GSDIM Super-Resolution Images Do you know that most protists and bacteria lack in one feature that each of our body cell has? Our cells are touch and communicate with one another. They send and receive a variety of signals that coordinate their behavior to act together as a functional multicellular organism. Exploring the way of cellular communication and the ways how the cell surface interacts to organize tissues and body structures is of great interest. Kees Jalink and his team of scientists at the Netherlands Cancer Institute (NKI) in Amsterdam obtained new scientific insights into the molecular architecture of hemidesmosomes, cytoskeletal components, cell surface receptors and vesicular proteins with the help of Ground-State-Depletion (GSD)/ dSTORM microscopy. In this interview, Kees Jalink comments on their developments in imaging chambers, buffer conditions and image analysis to get the perfect super resolution image. https://www.leica-microsystems.com//science-lab/practical-guide-for-excellent-gsdim-super-resolution-images/ Wed, 26 Oct 2016 06:58:00 +0000 PhD Kees Jalink, PhD Tamara Straube, MSc Leila Nahidiazar, MSc Daniela Leyton Puig https://www.leica-microsystems.com/18149 EM Sample Preparation Structural Study of C. elegans Application Note for Leica EM ICE, Leica EM AFS2 - Wildtype L4 stage C. elegans (N2 strain) were placed in the 100 μm deep side of Lecithin-coated (see detailed protocol*) type A 3 mm Cu/Au carriers (Leica) with extracellular filler containing 1% (w/v) Agarose type IX and 2% (w/v) Bovine Serum Albumin in bacteria medium (see preparation details**) and sandwiched with the flat side of Lecithin-coated type B 3 mm Cu/Au carriers (Leica). Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/structural-study-of-c-elegans/ Fri, 16 Sep 2016 08:15:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. Rob Mesman https://www.leica-microsystems.com/18301 EM Sample Preparation Freeze-Fracture Replication of Pyramidal Cells Application Note for Leica EM HPM100 - Frozen samples (90 μm thick slices frozen by HPM100) were inserted into a double replica table and then fractured into two pieces at –130°C (after insertion of the tissue into BAF 060 the samples should be left in the chamber for 20 min to reach the –130°C). https://www.leica-microsystems.com//science-lab/freeze-fracture-replication-of-pyramidal-cells/ Thu, 08 Sep 2016 16:23:00 +0000 Akos Kulik https://www.leica-microsystems.com/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/18102 Super-Resolution The Actin Cytoskeleton Modulates the Activation of iNKT Cells by Segregating CD1d Nanoclusters on Antigen-Presenting Cells The ability of invariant natural killer T (iNKT) cells to recognize endogenous antigens represents a distinct immune recognition strategy, which underscores the constitutive memory phenotype of iNKT cells and their activation during inflammatory conditions. By using superresolution microscopy, we show that CD1d molecules form nanoclusters at the cell surface of APCs, and their size and density are constrained by the actin cytoskeleton. https://www.leica-microsystems.com//science-lab/the-actin-cytoskeleton-modulates-the-activation-of-inkt-cells-by-segregating-cd1d-nanoclusters-on-antigen-presenting-cells/ Wed, 10 Aug 2016 13:28:00 +0000 https://www.leica-microsystems.com/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/18164 Super-Resolution TFG Promotes Organization of Transitional ER and Efficient Collagen Secretion Collagen is the most abundant protein in the animal kingdom. It is of fundamental importance during development for cell differentiation and tissue morphogenesis as well as in pathological processes such as fibrosis and cancer cell migration. However, our understanding of the mechanisms of procollagen secretion remains limited. Here, we show that TFG organizes transitional ER (tER) and ER exit sites (ERESs) into larger structures. Depletion of TFG results in dispersion of tER elements that remain associated with individual ER-Golgi intermediate compartments (ERGICs) as largely functional ERESs. We show that TFG is not required for the transport and packaging of small soluble cargoes but is necessary for the export of procollagen from the ER. Our work therefore suggests a key relationship between the structure and function of ERESs and a central role for TFG in optimizing COPII assembly for procollagen export. https://www.leica-microsystems.com//science-lab/tfg-promotes-organization-of-transitional-er-and-efficient-collagen-secretion/ Tue, 14 Jun 2016 08:36:00 +0000 https://www.leica-microsystems.com/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/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/16565 Super-Resolution Cross-strand Binding of TFAM to a Single mtDNA Molecule Forms the Mitochondrial Nucleoid Scientists from three Max Planck Institutes have gained fundamental insights into the organization of mitochondrial DNA (mtDNA). The researchers observed in high-resolution images gained with nobel prize-winning microscopy techniques that single copies of mtDNA are packaged by a specialized protein into slightly elongated structures of circa 100 nm in length. https://www.leica-microsystems.com//science-lab/cross-strand-binding-of-tfam-to-a-single-mtdna-molecule-forms-the-mitochondrial-nucleoid/ Tue, 17 Nov 2015 17:20:00 +0000 Dr. Christian Kukat https://www.leica-microsystems.com/13935 Fluorescence Microscopy Video Talk by Roger Tsien: Fluorescent Proteins Live cell imaging has been revolutionized by the discovery of the green fluorescent protein (GFP). This lecture covers the history of GFP, how it folds and becomes fluorescent, how it has been mutated to produce additional colors (blue, cyan, yellow), and the discovery of red fluorescent proteins from corals. It also covers novel photoswitchable and photoactivatible fluorescent proteins, whose color can be changed by light, and new infrared fluorescent proteins. https://www.leica-microsystems.com//science-lab/video-talk-by-roger-tsien-fluorescent-proteins/ Wed, 07 Oct 2015 16:55:00 +0000 PhD Roger Y. Tsien https://www.leica-microsystems.com/16035 Fluorescence Microscopy Live-Cell Imaging Basics in Microscopy Fluorescent Proteins Illuminate Cell Biology Green fluorescent protein (GFP) isolated from the jellyfish Aequorea victoria and GFP-like fluorescent proteins from other animals have had an important role in the technical innovations that have driven these advances. This poster provides a comprehensive user's guide to fluorescent proteins and sensors , their key properties and the cell biological questions to which they can be applied. https://www.leica-microsystems.com//science-lab/fluorescent-proteins-illuminate-cell-biology/ Fri, 18 Sep 2015 09:03:00 +0000 https://www.leica-microsystems.com/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/14400 Basics in Microscopy Video Talk by Joseph Gall: Early History of Microscopy Joseph Gall takes us through the history of early microscopes and the discovery of the cell. Compound microscopes were invented alongside the telescope in the 17th century; however these microscopes were not widely used until the late 19th century due to optical aberrations. In the meantime, simple microscopes were used throughout the 1700s and 1800s to make major discoveries in biology, including the first descriptions of the nucleus, cilia, cells, bacteria, and protozoans. Once optics improved in the mid to late 1800s, compound microscopes were used to discover chromosomes, mitosis, and other cellular structures. https://www.leica-microsystems.com//science-lab/video-talk-by-joseph-gall-early-history-of-microscopy/ Tue, 11 Aug 2015 07:48:00 +0000 BS, PhD Joseph Gall https://www.leica-microsystems.com/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/16019 Super-Resolution Abstracts of the 5th European Super-Resolution User-Club Meeting The 5th Super-Resolution User Club Meeting was held in collaboration with Professor Kees Jalink and The Netherlands Cancer Institute (NKI) in Amsterdam. Having the meeting at a location where super-resolution microscopy is used on a daily basis makes a big difference, offering participants the chance to use live cells for workshops and see systems working in their true environments. Thanks also to the scientists that supported the meeting by coming and giving talks. As super-resolution continues to grow in importance in research, we recognize the need to come together to network, share information and experiences. Here we present the abstracts of the talks. https://www.leica-microsystems.com//science-lab/abstracts-of-the-5th-european-super-resolution-user-club-meeting/ Tue, 23 Jun 2015 13:23:00 +0000 PhD Kees Jalink, PhD Jürgen J. Schmied, PhD Jonas Ries, PhD David Williamson, MSc Daniela Leyton Puig, Robert Nieuwenhuizen, MSc Leila Nahidiazar, PhD Luc Reymond, PhD Imre Gaspar, Luca Lanzanò, Ivan Michel Antolovic https://www.leica-microsystems.com/14920 Multiphoton Microscopy CLEM Live-Cell Imaging Correlating Intravital Multi-Photon Microscopy to 3D Electron Microscopy of Invading Tumor Cells Using Anatomical Reference Points Cancer research unsing multiphoton microscopy and 3D electron microscopy. Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient events at high resolution. Performing correlative microscopy in complex and bulky samples such as an entire living organism is a time-consuming and error-prone task. https://www.leica-microsystems.com//science-lab/correlating-intravital-multi-photon-microscopy-to-3d-electron-microscopy-of-invading-tumor-cells-using-anatomical-reference-points/ Fri, 24 Apr 2015 13:06:00 +0000 https://www.leica-microsystems.com/15104 EM Sample Preparation Immersion Freezing for Cryo-Transmission Electron Microscopy: Applications A well established usage case for cryo-TEM is three-dimensional reconstruction of isolated macromolecules, virus particles, or filaments. On one hand, these approaches are based on averaging of repetitive structures – either due to numerous identical molecules, repetitive patterns on a filament, or symmetries, to reduce the noise inherent to cryo-TEM. https://www.leica-microsystems.com//science-lab/immersion-freezing-for-cryo-transmission-electron-microscopy-applications/ Mon, 09 Feb 2015 14:51:00 +0000 Dr. Guenter Resch https://www.leica-microsystems.com/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 Neuroscience Live-Cell Imaging Fluorescence Microscopy Confocal 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/14955 Super-Resolution Video Interview with Jean-Luc Vonesch Jean-Luc Vonesch is head of the imaging facility at the Institute of Genetics and Molecular and Cellular Biology (IGBMC), Strasburg, France. 23 years ago he was the founder of this facility which nowadays serves more than 850 scientists distributed among 47 working groups. Looking deeply into the cells is of a special interest Vonesch states. And with super-resolution microscopy he pretends it is easier to identify the regions of interest for subsequent electron microscopy: “And so we can gain time thanks to the super-resolution” he says. https://www.leica-microsystems.com//science-lab/video-interview-with-jean-luc-vonesch/ Wed, 31 Dec 2014 11:27:00 +0000 Dr. Jean-Luc Vonesch, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/14656 Confocal Microscopy Super-Resolution CLEM Live-Cell Imaging Quantitative Imaging ICln: A New Regulator of Non-Erythroid 4.1R Localisation and Function To optimise the efficiency of cell machinery, cells can use the same protein (often called a hub protein) to participate in different cell functions by simply changing its target molecules. There are large data sets describing protein-protein interactions ("interactome") but they frequently fail to consider the functional significance of the interactions themselves. https://www.leica-microsystems.com//science-lab/icln-a-new-regulator-of-non-erythroid-41r-localisation-and-function/ Mon, 03 Nov 2014 18:02:00 +0000 https://www.leica-microsystems.com/14406 EM Sample Preparation Brief Introduction to Freeze Fracture and Etching Freeze fracture describes the technique of breaking a frozen specimen to reveal internal structures. Freeze etching is the sublimation of surface ice under vacuum to reveal details of the fractured face that were originally hidden. A metal/carbon mix enables the sample to be imaged in a SEM (block-face) or TEM (replica). It is used to investigate for instance cell organelles, membranes, layers and emulsions. https://www.leica-microsystems.com//science-lab/brief-introduction-to-freeze-fracture-and-etching/ Wed, 01 Oct 2014 09:27:00 +0000 Gisela Höflinger https://www.leica-microsystems.com/14492 Fluorescence Microscopy Defect Transport Mechanism Leads to Shortened Chromosome Ends Scientists at the University of Göttingen have deciphered the biogenesis of an enzyme complex whose role is to ensure that the ends of chromosomes are not shortened during the cell division process and that the genetic material is fully maintained. The results were published in Cell Reports magazine. https://www.leica-microsystems.com//science-lab/defect-transport-mechanism-leads-to-shortened-chromosome-ends/ Fri, 26 Sep 2014 16:46:00 +0000 Haijia Wu https://www.leica-microsystems.com/14389 Super-Resolution TIRF Microscopy Video Interview with William Hughes William Hughes works at the Garvan Institute of Medical Research, Sydney (Australia). In his Lab Head position he is interested in the causes of diabetes particularly looking at changes in exocytic behavior of pancreatic beta cells as well as fat and muscle cells. TIRF microscopy is predestined for researchers looking at cellular processes near the cytoplasmic membrane. https://www.leica-microsystems.com//science-lab/video-interview-with-william-hughes/ Wed, 24 Sep 2014 15:14:00 +0000 Dr. William Hughes, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/13883 Super-Resolution Video Interview with Timo Zimmermann The first super-resolution image he saw was an eye opener for him: "It was not just structures that got smaller. I was looking at a sample that I specifically had high hopes of seeing another layer of complexity and this actually was there." https://www.leica-microsystems.com//science-lab/video-interview-with-timo-zimmermann/ Wed, 10 Sep 2014 09:19:00 +0000 Dr. Timo Zimmermann, Dipl. oec.-troph. Anja Schué, PhD Isabelle Köster https://www.leica-microsystems.com/14017 Super-Resolution Abstracts of the 4th European Super-Resolution User-Club Meeting The 4th Super-Resolution User Club Meeting was held in collaboration with Christian Eggeling and the Weatherall Institute of Molecular Medicine in Oxford, UK. Here we present the abstracts of the talks and interviews with participants. https://www.leica-microsystems.com//science-lab/abstracts-of-the-4th-european-super-resolution-user-club-meeting/ Fri, 08 Aug 2014 08:09:00 +0000 Prof. Christian Eggeling, Ph.D. Giuseppe Vicidomini, MSc Leila Nahidiazar, Dr. Sergi Padilla-Parra, Prof. Mark Neil, Dr. Marko Lampe, PhD Kees Jalink, Dr. Katrin Willig, Dr. Timo Zimmermann, Ph.D. Marc van Zandvoort https://www.leica-microsystems.com/14066 Multiphoton Microscopy Live-Cell Imaging 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/13839 Super-Resolution Super-resolution Molecular and Functional Imaging of Nanoscale Architectures in Life and Materials Science Super-resolution (SR) fluorescence microscopy has been revolutionizing the way in which we investigate the structures, dynamics, and functions of a wide range of nanoscale systems. In this review, I describe the current state of various SR fluorescence microscopy techniques along with the latest developments of fluorophores and labeling for the SR microscopy. https://www.leica-microsystems.com//science-lab/super-resolution-molecular-and-functional-imaging-of-nanoscale-architectures-in-life-and-materials-science/ Mon, 07 Jul 2014 11:45:00 +0000 https://www.leica-microsystems.com/13740 Super-Resolution Neuroscience Interview with Dr. Yasushi Okada Yasushi Okada, team leader at Riken Quantitative Biology Center in Osaka, Japan, investigates vesicular transport mechanisms in neuronal cells. As the size of transported vesicles is below 100 nm and the diameter of microtubules is about 25 nm, he uses super-resolution techniques to study the sophisticated machinery of neuronal transport. https://www.leica-microsystems.com//science-lab/interview-with-dr-yasushi-okada/ Tue, 03 Jun 2014 15:38:00 +0000 Dipl. oec.-troph. Anja Schué, MD, PhD Yasushi Okada, PhD Isabelle Köster https://www.leica-microsystems.com/13280 EM Sample Preparation Neuroscience Capturing Cellular Dynamics with Millisecond Temporal Resolution The combination of two powerful techniques: optogenetics and high-pressure freezing now makes it possible to visualize a dynamic cellular activity with temporal resolution of 5 milliseconds. By coupling a flash of light with high-pressure freezing, the process of vesicle recycling at the synapses can now be imaged by electron microscopy. https://www.leica-microsystems.com//science-lab/capturing-cellular-dynamics-with-millisecond-temporal-resolution/ Mon, 12 May 2014 13:16:00 +0000 PhD Shigeki Watanabe, PhD Erik M. Jørgensen https://www.leica-microsystems.com/13034 Confocal Microscopy Establishment and Characterization of an Omasal Epithelial Cell Model Derived from Dairy Calves for the Study of Small Peptide Absorption The objective of this study was to establish a primary culture of omasal epithelial cells (OECs) derived from dairy calves and to characterize its function in small peptide absorption. Bovine omasal tissues were obtained from newborn Chinese Holstein calves and digested with a 2.5% trypsin solution to obtain OECs. https://www.leica-microsystems.com//science-lab/establishment-and-characterization-of-an-omasal-epithelial-cell-model-derived-from-dairy-calves-for-the-study-of-small-peptide-absorption/ Mon, 05 May 2014 16:46:00 +0000 https://www.leica-microsystems.com/13257 Multiphoton Microscopy Live-Cell Imaging Tracking Glomerular Fate Over Long Time Distances Multi-channel multiphoton microscopy with dedicated optics for CLARITY. The glomerular filtration barrier (GFB) is a complex spatial structure within the kidney glomerulis where ultrafiltration takes place. Podocytes are critical elements of the GFB and take part in the filtration process. They have been shown to be involved in the development of kidney diseases. However, due to technical limitations, the mechanism of glomerular pathology is not well understood. https://www.leica-microsystems.com//science-lab/tracking-glomerular-fate-over-long-time-distances/ Mon, 14 Apr 2014 12:32:00 +0000 M.D., Ph.D. Janos Peti-Peterdi, PhD Isabelle Köster https://www.leica-microsystems.com/12418 Confocal Microscopy Live-Cell Imaging Glucose-Stimulated Calcium Dynamics in Islets of Langerhans in Acute Mouse Pancreas Tissue Slices In endocrine cells within islets of Langerhans calcium ions couple cell stimulation to hormone secretion. Since the advent of modern fluorimetry, numerous in vitro studies employing primarily isolated mouse islets have investigated the effects of various secretagogues on cytoplasmic calcium, predominantly in insulin-secreting beta cells. Due to technical limitations, insights of these studies are inherently limited to a rather small subpopulation of outermost cells. https://www.leica-microsystems.com//science-lab/glucose-stimulated-calcium-dynamics-in-islets-of-langerhans-in-acute-mouse-pancreas-tissue-slices/ Tue, 11 Mar 2014 07:00:00 +0000 https://www.leica-microsystems.com/12420 Confocal Microscopy Live-Cell Imaging 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/12426 Confocal Microscopy Neuroscience Cellular Organization of Cortical Barrel Columns is Whisker-specific The cellular organization of the cortex is of fundamental importance for elucidating the structural principles that underlie its functions. It has been suggested that reconstructing the structure and synaptic wiring of the elementary functional building block of mammalian cortices, the cortical column, might suffice to reverse engineer and simulate the functions of entire cortices. https://www.leica-microsystems.com//science-lab/cellular-organization-of-cortical-barrel-columns-is-whisker-specific/ Fri, 14 Feb 2014 13:01:00 +0000 https://www.leica-microsystems.com/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/11802 Super-Resolution Super-Resolution Microscopy Gives New Insights into Nuclear Pore Complex Organization The Nuclear Pore Complex (NPC) is a large complex in the nuclear membrane, representing the gate to the eukaryotic genetic makeup. Because of this outstanding function the structure of the NPC is of great interest. Anna Szymborska, scientist at the EMBL in Heidelberg, comments on her resaerch results and the potential of Ground State Depletion microscopy (GSD) for protein complex analysis in the following interview. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-gives-new-insights-into-nuclear-pore-complex-organization/ Wed, 11 Dec 2013 17:53:00 +0000 Dr. Anna Szymborska, Dr. Jan Ellenberg, Dr. Christoph Greb https://www.leica-microsystems.com/10555 Super-Resolution Three-Dimensional Super-Resolution GSDIM Microscopy With the new 3D GSDIM technique structures like the Golgi and the microtubular network are resolved not only laterally, but also in a third dimension. The principle is based on the use of optical astigmatism to determine the accurate lateral and axial position of individual fluorochromes. https://www.leica-microsystems.com//science-lab/three-dimensional-super-resolution-gsdim-microscopy/ Wed, 04 Sep 2013 15:24:00 +0000 PhD Tamara Straube, Sebastian Bänfer, Prof. Ralf Jacob https://www.leica-microsystems.com/10447 Laser Microdissection Live-Cell Imaging EM Sample Preparation Confocal Microscopy 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/10135 Super-Resolution Widefield Microscopy Abstracts of the 3rd European Super-Resolution User-Club Meeting The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and widefield super-resolution users from ten European countries took three days’ out to deepen their knowledge on super-resolution techniques and applications and make use of an opportunity for full exchange of experiences. https://www.leica-microsystems.com//science-lab/abstracts-of-the-3rd-european-super-resolution-user-club-meeting/ Mon, 08 Jul 2013 10:27:00 +0000 Prof. Alberto Diaspro, Ph.D. Illaria Testa, Prof. Silvio Rizzoli, Ph.D. Giuseppe Vicidomini, Prof. Ralf Jacob, Ph.D. Eric Hosy, Prof. Colin Sheppard, Zeno Lavagnino https://www.leica-microsystems.com/10106 Confocal Microscopy Stereo Microscopy Fluorescence Microscopy Organ Regeneration: An Unlikely Fish Tale Spectacular discoveries in cardiac tissue regeneration are rapidly moving researchers closer to the goal of harnessing regenerative techniques to repair the human heart. Only eleven years ago, Dr. Kenneth Poss, Professor of Cell Biology at Duke University and an Early Career Scientist of the Howard Hughes Medical Institute, published the first research to clearly visualize an example of cardiac tissue regeneration using fluorescence microscopy. https://www.leica-microsystems.com//science-lab/organ-regeneration-an-unlikely-fish-tale/ Thu, 20 Jun 2013 08:01:00 +0000 PhD Katharine H. Hendrix https://www.leica-microsystems.com/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/7735 Super-Resolution Widefield Microscopy Abstracts of the 2nd European Super-Resolution User-Club Meeting The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. With a mixture of engaging talks by key experts in the field of super-resolution microscopy and stimulating discussion sessions, the meeting proved as popular as last year’s event, attracting a wide range of scientists interested in both confocal and widefield super-resolution and sample preparation techniques. https://www.leica-microsystems.com//science-lab/abstracts-of-the-2nd-european-super-resolution-user-club-meeting/ Thu, 01 Nov 2012 23:00:00 +0000 Prof. Hjalmar Brismar, Prof. Dr. Dr. h.c. Stefan Hell, Prof. Christian Eggeling, Dr. Anna Szymborska, Prof. Alberto Diaspro, Ph.D. Eric Hosy, Ph.D. Giuseppe Vicidomini, Hans van der Voort https://www.leica-microsystems.com/7364 Confocal Microscopy Live-Cell Imaging Nobel Prize 2012 in Chemistry for Studies of G-Protein-Coupled Receptors The Royal Swedish Academy of Sciences has decided to award the Nobel Prize in Chemistry for 2012 to Robert J. Lefkowitz and Brian K. Kobilka for studies of G-protein–coupled receptors. https://www.leica-microsystems.com//science-lab/nobel-prize-2012-in-chemistry-for-studies-of-g-protein-coupled-receptors/ Thu, 11 Oct 2012 22:00:00 +0000 Nobel Media AB 2012 https://www.leica-microsystems.com/6792 Live-Cell Imaging Stereo Microscopy Taking the Long View In exploring how embryos take shape, John Wallingford has identified a key pathway involved in vertebrate development – and human disease. https://www.leica-microsystems.com//science-lab/taking-the-long-view/ Mon, 10 Sep 2012 22:00:00 +0000 https://www.leica-microsystems.com/5929 Multiphoton Microscopy Confocal Microscopy Order versus Disorder In modern biomaterial design the generation of an environment mimicking some of the extracellular matrix features is envisaged to support molecular cross-talk between cells and scaffolds during tissue formation/remodeling. In bone substitutes chemical biomimesis has been particularly exploited; conversely, the relevance of pre-determined scaffold architecture for regenerated bone outputs is still unclear. https://www.leica-microsystems.com//science-lab/order-versus-disorder/ Thu, 19 Apr 2012 22:00:00 +0000 Paolo Giannoni, Paolo Bianco, Prof. Alberto Diaspro, Anna Tampieri, Ugo Valbusa, Giuseppe Firpo, Roberto Marotta, Monica Sandri, Paolo Bianchini, Rodolfo Quarto https://www.leica-microsystems.com/5175 TIRF Microscopy Live-Cell Imaging Protein Transport Processes at the Apical Membrane of Polarized Epithelial Cells Due to their special role in organ function and the exchange of biological components some body cells developed certain polarization characteristics. These are reflected in differences of their plasma membrane composition. The essential and fascinating task of polarized protein transport in epithelial cells is to get the right protein into the right membrane. https://www.leica-microsystems.com//science-lab/protein-transport-processes-at-the-apical-membrane-of-polarized-epithelial-cells/ Sun, 11 Mar 2012 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5275 Confocal Microscopy How to Study Protein Recruitment to DNA Lesions by a Combination of UV Laser and White Light Laser Understanding how DNA lesions are optimally repaired is of functional significance, especially from the view of genome karyotype stability. https://www.leica-microsystems.com//science-lab/how-to-study-protein-recruitment-to-dna-lesions-by-a-combination-of-uv-laser-and-white-light-laser/ Wed, 22 Feb 2012 23:00:00 +0000 Assoc. Prof. Dr. Eva Bártová, Assoc. Prof. Dr. Stanislav Kozubek, Ph.D. Gabriela Sustáčková https://www.leica-microsystems.com/5217 Fluorescence Microscopy Live-Cell Imaging Neuroscience TIRF Microscopy Confocal 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/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/9689 Neuroscience Confocal Microscopy Organotypic Cerebellar Cultures: Apoptotic Challenges and Detection Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 and have constituted a major breakthrough in the field of neuroscience. Since then, the technique was developed further and currently there are many different ways to prepare organotypic cultures. The method presented here was adapted from the one described by Stoppini et al. for the preparation of the slices and from Gogolla et al. for the staining procedure. https://www.leica-microsystems.com//science-lab/organotypic-cerebellar-cultures-apoptotic-challenges-and-detection/ Tue, 17 May 2011 17:50:00 +0000 https://www.leica-microsystems.com/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/10319 CLEM EM Sample Preparation High Data Output and Automated 3D Correlative Light–Electron Microscopy Method Correlative light/electron microscopy (CLEM) allows the simultaneous observation of a given subcellular structure by fluorescence light microscopy (FLM) and electron microscopy. The use of this approach is becoming increasingly frequent in cell biology. In this study, we report on a new high data output CLEM method based on the use of cryosections. https://www.leica-microsystems.com//science-lab/high-data-output-and-automated-3d-correlative-light-electron-microscopy-method/ Sat, 15 Nov 2008 12:52:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/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