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  • 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.
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  • 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.
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  • Webinar: Introduction to Correlative Light and Electron Microscopy (CLEM)

    The webinar will provide an overview of the latest advances in Cryo CLEM, which acts as a powerful interface by combining the best of the light and electron microscopy worlds to overcome their independent barriers and determine the location of fluorescent labelled structures within the landscape of an electron micrograph and showcase how Cryo CLEM adds additional value to quantitative 3D imaging and tomography.
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  • 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.
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  • Webinar: Correlative Fluorescence and (Cryo-) Electron Microscopy

    In this webinar, you will learn how you can identify objects of interest based on their fluorescent signal, and then find and image them in 3D in the transmission electron microscope using correlative light and electron microscopy (CLEM) methods.
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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  • Webinar: Correlative Light Electron Microscopy: 1 + 1 = 3

    Imaging is one of the key technologies for Biomedical research. In many cases it is useful to obtain data from different imaging modalities. Especially combining 2 techniques in one single experiment, so-called Correlative Microscopy, can and should give a better answer than each technique alone. 1 + 1 = 3.
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  • 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.
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  • 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).
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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  • 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.
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