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Cryo-TEM

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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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  • Electron Microscopy Sample Preparation: “The Future is Cold, Dynamic and Hybrid”

    In 2014, the renowned Electron Microscopy for Materials Science (EMAT) research lab at the University Antwerp, Belgium, and Leica Microsystems started a fruitful collaboration to establish a Leica Reference Site in Antwerp. This site, officially opened in July 2014, is dedicated to specimen preparation for electron microscopy in materials science with a special focus on ion beam milling and recently also on carbon coating. In this interview Prof Gustaf van Tendeloo, Director of EMAT, and Frédéric Leroux, TEM specimen preparation specialist, talk about research topics at EMAT, how the Leica reference site has evolved, and future trends for EM sample preparation.
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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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  • 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.
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  • Immersion Freezing for Cryo-Transmission Electron Microscopy: Fundamentals

    The high vacuum required in a transmission electron microscope (TEM) greatly impairs the ability to study specimens naturally occurring in an aqueous phase: exposing "wet" specimens to a pressure significantly lower than the vapour pressure of water will lead to the water phase boiling off rapidly in the column, with devastating consequences for the structure of the specimen. Hence, various methods to dry specimens before inspection are employed in conventional TEM – a preparative step often associated with artifacts limiting the significance of the results.
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  • Immersion Freezing of Suspended Particles and Cells for Cryo-Electron Microscopy

    Immersion freezing of thin aqueous specimens is an essential preparation technique for cryo-transmission electron microscopy (cryo-TEM), aiming to preserve fragile biological structures such as molecules and cells in their hydrated environment for a close-to-native visualization. For successful experiments, vitreous ice must be produced, surface contamination must be avoided, and, most important, the natural state of the structure must be preserved.
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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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