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An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma. / Woud, Wouter W.; van der Pol, Edwin; Mul, Erik; Hoogduijn, Martin J.; Baan, Carla C.; Boer, Karin; Merino, Ana.

In: Communications Biology, Vol. 5, No. 1, 633, 01.12.2022.

Research output: Contribution to journalArticleAcademicpeer-review

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APA

Woud, W. W., van der Pol, E., Mul, E., Hoogduijn, M. J., Baan, C. C., Boer, K., & Merino, A. (2022). An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma. Communications Biology, 5(1), [633]. https://doi.org/10.1038/s42003-022-03569-5

Vancouver

Author

Woud, Wouter W. ; van der Pol, Edwin ; Mul, Erik ; Hoogduijn, Martin J. ; Baan, Carla C. ; Boer, Karin ; Merino, Ana. / An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma. In: Communications Biology. 2022 ; Vol. 5, No. 1.

BibTeX

@article{6bb5ebd97c4f44bf91be430cb6631363,
title = "An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma",
abstract = "Extracellular vesicles (EVs) are tissue-specific particles released by cells containing valuable diagnostic information in the form of various biomolecules. To rule out selection bias or introduction of artefacts caused by EV isolation techniques, we present a clinically feasible, imaging flow cytometry (IFCM)–based methodology to phenotype and determine the concentration of EVs with a diameter ≤400 nm in human platelet-poor plasma (PPP) without prior isolation of EVs. Instrument calibration (both size and fluorescence) were performed with commercial polystyrene beads. Detergent treatment of EVs was performed to discriminate true vesicular events from artefacts. Using a combination of markers (CFSE & Tetraspanins, or CD9 & CD31) we found that >90% of double-positive fluorescent events represented single EVs. Through this work, we provide a framework that will allow the application of IFCM for EV analysis in peripheral blood plasma in a plethora of experimental and potentially diagnostic settings. Additionally, this direct approach for EV analysis will enable researchers to explore corners of EVs as cellular messengers in healthy and pathological conditions.",
author = "Woud, {Wouter W.} and {van der Pol}, Edwin and Erik Mul and Hoogduijn, {Martin J.} and Baan, {Carla C.} and Karin Boer and Ana Merino",
note = "Funding Information: The authors would like to thank Peter Rhein (Luminex) for his support and discussions throughout the project. Publisher Copyright: {\textcopyright} 2022, The Author(s).",
year = "2022",
month = dec,
day = "1",
doi = "10.1038/s42003-022-03569-5",
language = "English",
volume = "5",
journal = "Communications Biology",
issn = "2399-3642",
publisher = "Springer Nature",
number = "1",

}

RIS

TY - JOUR

T1 - An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma

AU - Woud, Wouter W.

AU - van der Pol, Edwin

AU - Mul, Erik

AU - Hoogduijn, Martin J.

AU - Baan, Carla C.

AU - Boer, Karin

AU - Merino, Ana

N1 - Funding Information: The authors would like to thank Peter Rhein (Luminex) for his support and discussions throughout the project. Publisher Copyright: © 2022, The Author(s).

PY - 2022/12/1

Y1 - 2022/12/1

N2 - Extracellular vesicles (EVs) are tissue-specific particles released by cells containing valuable diagnostic information in the form of various biomolecules. To rule out selection bias or introduction of artefacts caused by EV isolation techniques, we present a clinically feasible, imaging flow cytometry (IFCM)–based methodology to phenotype and determine the concentration of EVs with a diameter ≤400 nm in human platelet-poor plasma (PPP) without prior isolation of EVs. Instrument calibration (both size and fluorescence) were performed with commercial polystyrene beads. Detergent treatment of EVs was performed to discriminate true vesicular events from artefacts. Using a combination of markers (CFSE & Tetraspanins, or CD9 & CD31) we found that >90% of double-positive fluorescent events represented single EVs. Through this work, we provide a framework that will allow the application of IFCM for EV analysis in peripheral blood plasma in a plethora of experimental and potentially diagnostic settings. Additionally, this direct approach for EV analysis will enable researchers to explore corners of EVs as cellular messengers in healthy and pathological conditions.

AB - Extracellular vesicles (EVs) are tissue-specific particles released by cells containing valuable diagnostic information in the form of various biomolecules. To rule out selection bias or introduction of artefacts caused by EV isolation techniques, we present a clinically feasible, imaging flow cytometry (IFCM)–based methodology to phenotype and determine the concentration of EVs with a diameter ≤400 nm in human platelet-poor plasma (PPP) without prior isolation of EVs. Instrument calibration (both size and fluorescence) were performed with commercial polystyrene beads. Detergent treatment of EVs was performed to discriminate true vesicular events from artefacts. Using a combination of markers (CFSE & Tetraspanins, or CD9 & CD31) we found that >90% of double-positive fluorescent events represented single EVs. Through this work, we provide a framework that will allow the application of IFCM for EV analysis in peripheral blood plasma in a plethora of experimental and potentially diagnostic settings. Additionally, this direct approach for EV analysis will enable researchers to explore corners of EVs as cellular messengers in healthy and pathological conditions.

UR - http://www.scopus.com/inward/record.url?scp=85132970990&partnerID=8YFLogxK

U2 - 10.1038/s42003-022-03569-5

DO - 10.1038/s42003-022-03569-5

M3 - Article

C2 - 35768629

VL - 5

JO - Communications Biology

JF - Communications Biology

SN - 2399-3642

IS - 1

M1 - 633

ER -

ID: 24873980