A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry

Author Identifier

Michael Edward Clark
ORCID: 0000-0002-9748-5221

Elin Gray
ORCID: 0000-0002-8613-3570

Document Type

Journal Article

Publication Title

Journal of Immunological Methods

ISSN

1872-7905

Volume

468

First Page

61

Last Page

66

PubMed ID

30885719

Publisher

Elsevier BV

School

School of Medical and Health Sciences

RAS ID

28793

Comments

Armitage, J. D., Tan, D. B. A., Cha, L., Clark, M., Gray, E. S., Fuller, K. A., & Moodley, Y. P. (2019). A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry. Journal of immunological methods, 468, 61-66. Available here

Abstract

Flow cytometry provides robust, multi-parametric and quantitative information on single cells which also exhibits enormous potential as a tool for small particle characterisation. Small extracellular vesicle (sEV) detection by flow cytometry remains compromised due to the high prevalence of swarm detection, which is defined by the simultaneous illumination of more than one sEV, recorded as a single event. Detection of sEVs by imaging flow cytometry presents a major advantage by having the ability to resolve single particles from swarm detection based on the image features recorded for each event. In this study, we provide a simplified protocol that facilitates the removal of both swarm events and aggregated particles to improve the accuracy of sEV analysis. Our results indicate that imaging flow cytometry should be at the forefront as a robust and sensitive technique for sEV characterisation.

DOI

10.1016/j.jim.2019.03.006

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