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

J. D. Armitage, D. B.A. Tan, L. Cha, M. Clark, E. S. Gray, K. A. Fuller, Y. P. Moodley

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)61-66
Number of pages6
JournalJournal of Immunological Methods
Volume468
DOIs
Publication statusPublished - 1 May 2019

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Flow Cytometry
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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.",
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A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry. / Armitage, J. D.; Tan, D. B.A.; Cha, L.; Clark, M.; Gray, E. S.; Fuller, K. A.; Moodley, Y. P.

In: Journal of Immunological Methods, Vol. 468, 01.05.2019, p. 61-66.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry

AU - Armitage, J. D.

AU - Tan, D. B.A.

AU - Cha, L.

AU - Clark, M.

AU - Gray, E. S.

AU - Fuller, K. A.

AU - Moodley, Y. P.

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AB - 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.

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KW - Immunophenotyping

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