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
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
Access Rights
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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