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Characterization of Extracellular Vesicles by Transmission Electron Microscopy and Immunolabeling Electron Microscopy

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Cell-Secreted Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2668))

Abstract

Transmission electron microscopy (TEM) is currently the only method that enables the observation of extracellular vesicles (EVs) at a nanometer scale. Direct visualization of the whole content of EV preparation provides not only crucial insights on the morphology of EVs but also an objective evaluation of the content and purity of the preparation. Coupled to immunogold labeling, TEM allows the detection and association of proteins at the surface of EVs. In these techniques, EVs are deposited on grids and are chemically immobilized and contrasted to withstand a high-voltage electron beam. Under high vacuum, the electron beam hits the sample and the electrons that scatter forward are collected to form an image. Here, we describe the steps needed to observe EVs by classical TEM and the extra steps required to label proteins through immunolabeling electron microscopy (IEM).

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Acknowledgments

We would like to thank Roberta Palmulli to provide electron microscopy pictures and Maryse Romao for the pictures of grids handling. MLC is financed by ANR ZENITH (ANR-20-CE18-0026_01). GVN is supported by the fondation ARC and Institut National du Cancer-projets libres “Biologie et Sciences du Cancer”’ (INCa-PLBIO2019-125).

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Authors and Affiliations

  1. Université de Paris, Institute of Psychiatry and Neuroscience of Paris (IPNP), Paris, France

    Maribel Lara Corona

  2. Institut Curie, PSL Research University, CNRS, UMR144, Cell and Tissue Imaging Facility(PICT-IBiSA), Paris, France

    Ilse Hurbain, Graça Raposo & Guillaume van Niel

  3. GHU Paris Psychiarie et Neurosciences, Hôpital Sainte Anne, Paris, France

    Guillaume van Niel

Authors
  1. Maribel Lara Corona
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  2. Ilse Hurbain
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  3. Graça Raposo
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  4. Guillaume van Niel
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Corresponding author

Correspondence to Guillaume van Niel .

Editor information

Editors and Affiliations

  1. GeneCellNano, Oulu University, Oulu, Finland

    Seppo Vainio

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Corona, M.L., Hurbain, I., Raposo, G., van Niel, G. (2023). Characterization of Extracellular Vesicles by Transmission Electron Microscopy and Immunolabeling Electron Microscopy. In: Vainio, S. (eds) Cell-Secreted Vesicles. Methods in Molecular Biology, vol 2668. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3203-1_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3203-1_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3202-4

  • Online ISBN: 978-1-0716-3203-1

  • eBook Packages: Springer Protocols

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