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. 2023:2668:33-43.
doi: 10.1007/978-1-0716-3203-1_4.

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

Maribel Lara Corona  1 , Ilse Hurbain  2 , Graça Raposo  2 , Guillaume van Niel  3   4
Affiliations
Free article

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

Maribel Lara Corona et al. Methods Mol Biol. 2023.
Free article

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).

Keywords: Antibodies (Ab); Extracellular vesicles (EVs); Immunogold labeling; Immunolabeling electron microscopy (IEM); Transmission electron microscopy (TEM).

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References

    1. Wolf P (1967) The nature and significance of platelet products in human plasma. Br J Haematol 13(3):269–288). Wiley. https://doi.org/10.1111/j.1365-2141.1967.tb08741.x - DOI - PubMed
    1. Harding C, Stahl P (1983) Transferrin recycling in reticulocytes: pH and iron are important determinants of ligand binding and processing. Biochem Biophys Res Commun 113(2):650–658). Elsevier BV. https://doi.org/10.1016/0006-291x(83)91776-x - DOI - PubMed
    1. Pan B-T, Johnstone RM (1983) Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro: selective externalization of the receptor. Cell 33(3):967–978). Elsevier BV. https://doi.org/10.1016/0092-8674(83)90040-5 - DOI - PubMed
    1. Colombo M, Moita C, van Niel G, Kowal J, Vigneron J, Benaroch P, Manel N, Moita LF, Théry C, Raposo G (2013) Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. J Cell Sci Comp Biolog. https://doi.org/10.1242/jcs.128868
    1. Jeppesen DK, Fenix AM, Franklin JL, Higginbotham JN, Zhang Q, Zimmerman LJ, Liebler DC, Ping J, Liu Q, Evans R, Fissell WH, Patton JG, Rome LH, Burnette DT, Coffey RJ (2019) Reassessment of exosome composition. Cell 177(2):428–445). Elsevier BV. https://doi.org/10.1016/j.cell.2019.02.029 - DOI - PubMed - PMC

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