Negative Staining of Purified Exosomes: A Procedure to Visualize Exosome Morphology Using Transmission Electron Microscopy

Published: April 30, 2023

Abstract

Source: Jung, M. K. et al. Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy. J. Vis. Exp. (2018)

This video describes a procedure for negative staining of purified exosomes. The negatively stained exosomes are imaged using transmission electron microscopy. This technique helps visualize the morphology of exosomes.

Protocol

1. Negative Staining Glow-discharge the thin formvar/carbon film-coated 200 mesh copper EM grids for 1 min by glow discharger. Fix purified exosomes with 1 mL of 2% paraformaldehyde (PFA) for 5 min. NOTE: Paraformaldehyde fumes are toxic. All work should be done in a ventilated fume hood. Load 5-7 µL exosome suspension solution on the grid and incubate for 1 min. If the concentration of exosome is too high, dilute the concentr…

Divulgaciones

The authors have nothing to disclose.

Materials

Uranyl acetate EMS 22400 Hazardous chemical
Transmission Electron Microscopy JEOL JEM2200FS
Glow discharger JEOL JFC1100E
Paraformaldehyde EMS 19210
Formvar carbon coated Copper
Grid (200 mesh)
EMS FCF200-CU
Formvar carbon coated Nickel
Grid (200 mesh)
EMS FCF200-NI

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Negative Staining of Purified Exosomes: A Procedure to Visualize Exosome Morphology Using Transmission Electron Microscopy. J. Vis. Exp. (Pending Publication), e20420, doi: (2023).

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