Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy

Exosomes are nano-sized extracellular vesicles secreted by body fluids and are known to represent the characteristics of cells that secrete them. The contents and morphology of the secreted vesicles reflect cell behavior or physiological status, for example cell growth, migration, cleavage, and deat...

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Bibliographic Details
Published in:Journal of visualized experiments no. 131
Main Authors: Jung, Min Kyo, Mun, Ji Young
Format: Journal Article
Language:English
Published: United States MyJove Corporation 04.01.2018
Subjects:
atomic force microscopy
body fluids
cell growth
Cell Proliferation - physiology
Cellular Biology
Colonic Neoplasms - chemistry
Colonic Neoplasms - ultrastructure
death
exosomes
Exosomes - chemistry
HCT116 Cells
Humans
image analysis
Immunohistochemistry - methods
Microscopy, Electron, Transmission - methods
Microtomy - methods
physiological state
scanning electron microscopy
staining
Staining and Labeling - methods
transmission electron microscopy
ISSN:1940-087X, 1940-087X
Online Access:Get full text
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Summary:Exosomes are nano-sized extracellular vesicles secreted by body fluids and are known to represent the characteristics of cells that secrete them. The contents and morphology of the secreted vesicles reflect cell behavior or physiological status, for example cell growth, migration, cleavage, and death. The exosomes' role may depend highly on size, and the size of exosomes varies from 30 to 300 nm. The most widely used method for exosome imaging is negative staining, while other results are based on Cryo-Transmission Electron Microscopy, Scanning Electron Microscopy, and Atomic Force Microscopy. The typical exosome's morphology assessed through negative staining is a cup-shape, but further details are not yet clear. An exosome well-characterized through structural study is necessary particular in medical and pharmaceutical fields. Therefore, function-dependent morphology should be verified by electron microscopy techniques such as labeling a specific protein in the detailed structure of exosome. To observe detailed structure, ultrathin sectioned images and negative stained images of exosomes were compared. In this protocol, we suggest transmission electron microscopy for the imaging of exosomes including negative staining, whole mount immuno-staining, block preparation, thin section, and immuno-gold labelling.
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Correspondence to: Ji Young Mun at mjy1026@gmail.com
ISSN:1940-087X
1940-087X
DOI:10.3791/56482