Leica Science Lab - Tag : CLEM https://www.leica-microsystems.com//science-lab/tag/tags/clem/show/Tag/ Article tagged with CLEM en-US https://www.leica-microsystems.com/25022 EM Sample Preparation Bridging Structure and Dynamics at the Nanoscale through Optogenetics and Electrical Stimulation Nanoscale ultrastructural information is typically obtained by means of static imaging of a fixed and processed specimen. However, this is only a snapshot of one moment within a dynamic system in which structures are constantly changing. Exploring specific time points of a dynamic process is therefore a major challenge. Exploring a process at the nanoscale through optogenetics or electrical field stimulation in combination with timed millisecond precision vitrification is a promising technology to overcome this challenge. In the first part of a series of application notes the practical considerations of stimulation-assisted vitrification are discussed. https://www.leica-microsystems.com//science-lab/bridging-structure-and-dynamics-at-the-nanoscale-through-optogenetics-and-electrical-stimulation/ Mon, 20 May 2019 22:00:00 +0000 Dr. Andres Kaech, PhD Frédéric Leroux https://www.leica-microsystems.com/19607 EM Sample Preparation Interview with Dr. Shigeki Watanabe on Research in Synaptic Membrane Dynamics Dr. Shigeki Watanabe, principle investigator of the department of Cell Biology at the Johns Hopkins University School of Medicine in Baltimore, held a workshop in Zürich, Switzerland on methods to study synaptic dynamics with millisecond precision. In collaboration with Dr. Andres Käch from the University of Zurich all workshop attendees enjoyed presentations and hands-on sessions on the EM ICE by Leica Microsystems with Light and Electrical Stimulation, revealing the latest developments in brain research. During this workshop Dr. Bernd Sägmüller from Leica Microsystems had the chance for an interview with Dr. Watanabe. https://www.leica-microsystems.com//science-lab/interview-with-dr-shigeki-watanabe-on-research-in-synaptic-membrane-dynamics/ Thu, 06 Jul 2017 23:00:00 +0000 Dr. Bernd Sägmüller, PhD Shigeki Watanabe https://www.leica-microsystems.com/18008 CLEM Fluorescence Microscopy EM Sample Preparation Correlative Cryo-Fluorescence and Cryo-Scanning Electron Microscopy as a Straightforward Tool to Study Host-Pathogen Interactions Correlative light and electron microscopy is an imaging technique that enables identification and targeting of fluorescently tagged structures with subsequent imaging at near-to-nanometer resolution. We established a novel correlative cryo-fluorescence microscopy and cryo-scanning electron microscopy workflow, which enables imaging of the studied object of interest very close to its natural state, devoid of artifacts caused for instance by slow chemical fixation. This system was tested by investigating the interaction of the zoonotic bacterium Borrelia burgdorferi with two mammalian cell lines of neural origin in order to broaden our knowledge about the cell-association mechanisms that precedes the entry of the bacteria into the cell. https://www.leica-microsystems.com//science-lab/correlative-cryo-fluorescence-and-cryo-scanning-electron-microscopy-as-a-straightforward-tool-to-study-host-pathogen-interactions/ Tue, 13 Dec 2016 07:58:00 +0000 Martin Strnad https://www.leica-microsystems.com/17382 CLEM Imaging of Host Cell-bacteria Interactions using Correlative Microscopy under Cryo-conditions Pathogenic bacteria have developed intriguing strategies to establish and promote infections in their respective hosts. Most bacterial pathogens initiate infectious diseases by adhering to host cells surface. Knowledge of the interplay between the pathogenic organism and the host cells can provide fundamental new insight into the underlying mechanisms of the infectious process and therefore of the disease. Various microscopy techniques have proven to be critical tools to study these events. https://www.leica-microsystems.com//science-lab/imaging-of-host-cell-bacteria-interactions-using-correlative-microscopy-under-cryo-conditions/ Tue, 01 Mar 2016 11:34:00 +0000 Martin Strnad https://www.leica-microsystems.com/15509 CLEM Correlative In-Resin Super-Resolution and Electron Microscopy Using Standard Fluorescent Proteins We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer superior structural preservation, performing in-resin super-resolution, however, remains a challenge. https://www.leica-microsystems.com//science-lab/correlative-in-resin-super-resolution-and-electron-microscopy-using-standard-fluorescent-proteins/ Thu, 04 Jun 2015 10:28:00 +0000 https://www.leica-microsystems.com/15106 EM Sample Preparation Fluorescence Microscopy Cryo CLEM – the Combination of Cryo Fluorescence Microscopy with Cryo Electron Microscopy Many biological insights can be obtained by combining the power of Fluorescence Microscopy (FM) with that of Electron Microscopy (EM) to study the same sample – this is called Correlative Light and Electron Microscopy (CLEM). In FM, specific proteins can be labelled and identified, and their dynamics and interactions can be visualized in fixed or living cells. https://www.leica-microsystems.com//science-lab/cryo-clem-the-combination-of-cryo-fluorescence-microscopy-with-cryo-electron-microscopy/ Wed, 18 Mar 2015 08:17:00 +0000 PhD Martin Schorb, PhD John A. G. Briggs https://www.leica-microsystems.com/14828 Laser Microdissection CLEM Confocal Microscopy Live-Cell Imaging Customized Patterned Substrates for Highly Versatile Correlative Light-Scanning Electron Microscopy Correlative light electron microscopy (CLEM) combines the advantages of light and electron microscopy, thus making it possible to follow dynamic events in living cells at nanometre resolution. Various CLEM approaches and devices have been developed, each of which has its own advantages and technical challenges. We here describe our customized patterned glass substrates, which improve the feasibility of correlative fluorescence/confocal and scanning electron microscopy. https://www.leica-microsystems.com//science-lab/customized-patterned-substrates-for-highly-versatile-correlative-light-scanning-electron-microscopy/ Fri, 06 Mar 2015 17:14:00 +0000 https://www.leica-microsystems.com/15081 EM Sample Preparation CLEM New Insights into Cilia and Flagella by Cryo-EM Cilia and flagella were the first organelles to be discovered and have been studied for centuries. However, their essential role in humans and how ciliary defects cause diseases are still not well understood. Cryo-EM has recently shed new light on their inner workings and solved some long-standing mysteries, only to raise new questions on how cilia and flagella function. https://www.leica-microsystems.com//science-lab/new-insights-into-cilia-and-flagella-by-cryo-em/ Mon, 16 Feb 2015 09:25:00 +0000 PhD Thomas Heuser https://www.leica-microsystems.com/14887 EM Sample Preparation Video Tutorials: Filling and Assembling of Different Carriers for High-Pressure Freezing High pressure freezing (HPF) is a cryo-fixation method primarily for biological samples, but also for a variety of non-biological materials. It is a technique that yields optimal preservation in many cell types and tissues or in organic and inorganic composites. Most commonly, the high pressure frozen samples are analyzed further with light or electron microscopy after appropriate processing. https://www.leica-microsystems.com//science-lab/video-tutorials-filling-and-assembling-of-different-carriers-for-high-pressure-freezing/ Fri, 05 Dec 2014 08:45:00 +0000 Dr. Cveta Tomova https://www.leica-microsystems.com/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/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/10281 CLEM EM Sample Preparation Live-Cell Imaging Confocal Microscopy 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/10060 EM Sample Preparation CLEM Brief Introduction to High-Pressure Freezing Water is the most abundant cellular constituent and therefore important for preserving cellular ultra-structure. Currently the only way to fix cellular constituents without introducing significant structural alterations is by cryo-fixation. There are currently two common methods employed; plunge freezing and high pressure freezing. https://www.leica-microsystems.com//science-lab/brief-introduction-to-high-pressure-freezing/ Wed, 12 Jun 2013 17:59:00 +0000 Dr. Cveta Tomova https://www.leica-microsystems.com/10886 Live-Cell Imaging EM Sample Preparation CLEM Postlipolytic Insulin-dependent Remodeling of Micro Lipid Droplets in Adipocytes Despite the lipolysis–lipogenesis cycle being a fundamental process in adipocyte biology, very little is known about the morphological changes that occur during this process. The remodeling of lipid droplets to form micro lipid droplets (mLDs) is a striking feature of lipolysis in adipocytes, but once lipolysis ceases, the cell must regain its basal morphology. https://www.leica-microsystems.com//science-lab/postlipolytic-insulin-dependent-remodeling-of-micro-lipid-droplets-in-adipocytes/ Tue, 15 May 2012 14:14:00 +0000 https://www.leica-microsystems.com/10866 CLEM Fluorescence Microscopy EM Sample Preparation A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms Electron microscopy (EM) achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ visualization in cells and tissues. Here we introduce ‘"miniSOG"’ (for mini Singlet Oxygen Generator), a fluorescent flavoprotein engineered from Arabidopsis phototropin 2. https://www.leica-microsystems.com//science-lab/a-genetically-encoded-tag-for-correlated-light-and-electron-microscopy-of-intact-cells-tissues-and-organisms/ Sun, 04 Sep 2011 19:37:00 +0000 https://www.leica-microsystems.com/10831 CLEM EM Sample Preparation Advanced Correlative Light/Electron Microscopy: Current Methods and New Developments Using Tokuyasu Cryosections Microscopy is an essential tool for analysis of cellular structures and function. With the advent of new fluorescent probes and super-resolution light microscopy techniques, the study of dynamic processes in living cells has been greatly facilitated. Fluorescence light microscopy provides analytical, quantitative, and three-dimensional (3D) data with emphasis on analysis of live cells using fluorescent markers. https://www.leica-microsystems.com//science-lab/advanced-correlative-lightelectron-microscopy-current-methods-and-new-developments-using-tokuyasu-cryosections/ Sun, 20 Dec 2009 09:11:00 +0000 https://www.leica-microsystems.com/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/7970 CLEM EM Sample Preparation CLEM: Combining the Strengths of Light and Electron Microscopy In recent years light microscopy studies have been dominated by live cell imaging while electron microscopy has been used for high-resolution studies. Latterly, there has been increasing interest in combining these techniques. This combination is called Correlative Light Electron Microscopy (CLEM). Due to the high resolution made possible by electron microscopy, artefacts induced during preparation of a sample can, however, also be clearly seen. https://www.leica-microsystems.com//science-lab/clem-combining-the-strengths-of-light-and-electron-microscopy/ Mon, 09 Apr 2007 22:00:00 +0000 Dr. Paul Verkade