TY - JOUR
T1 - Standardization of extracellular vesicle concentration measurements by flow cytometry
T2 - the past, present, and future
AU - Bettin, Britta A.
AU - Varga, Zoltán
AU - Nieuwland, Rienk
AU - van der Pol, Edwin
N1 - Funding Information: Funding information B.B. acknowledges funding by METVES II. This project (METVES II) has received funding from the EMPIR program cofinanced by the Participating States and from the European Union's Horizon 2020 research and innovation program. E.v.d.P. acknowledges funding from the Netherlands Organisation for Scientific Research—Domain Applied and Engineering Sciences (NWO-TTW), research programs VENI 15924 and VIDI 19724. Z.V. was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences. Funding Information: Funding information B.B. acknowledges funding by METVES II. This project (METVES II) has received funding from the EMPIR program cofinanced by the Participating States and from the European Union’s Horizon 2020 research and innovation program. E.v.d.P. acknowledges funding from the Netherlands Organisation for Scientific Research—Domain Applied and Engineering Sciences (NWO-TTW), research programs VENI 15924 and VIDI 19724. Z.V. was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences . Publisher Copyright: © 2023 The Authors
PY - 2023/8
Y1 - 2023/8
N2 - Concentrations of extracellular vesicles (EVs) in body fluids are being explored as disease biomarkers. Most laboratories use flow cytometry to characterize single EVs at high throughput. A flow cytometer (FCM) detects light scattering and fluorescence intensities of EVs. However, detection of EVs by flow cytometry is complicated for 2 reasons. First, EVs are small and have weak light scattering and fluorescence signals compared to cells and are, therefore, hard to detect. Second, FCMs differ in sensitivity and provide data in arbitrary units, which complicates data interpretation. Due to the mentioned challenges, the measured concentration of EVs by flow cytometry is cumbersome to compare between FCMs and institutes. To improve comparability, standardization and development of traceable reference materials to calibrate all aspects of an FCM are needed, as are interlaboratory comparison studies. Within this article, we will provide an overview of the standardization of EV concentration measurements, including the current effort to introduce robust calibration of FCMs, thereby enabling comparable concentration measurements of EVs, which in turn can be used to establish clinically relevant reference ranges of EV concentrations in blood plasma and other body fluids.
AB - Concentrations of extracellular vesicles (EVs) in body fluids are being explored as disease biomarkers. Most laboratories use flow cytometry to characterize single EVs at high throughput. A flow cytometer (FCM) detects light scattering and fluorescence intensities of EVs. However, detection of EVs by flow cytometry is complicated for 2 reasons. First, EVs are small and have weak light scattering and fluorescence signals compared to cells and are, therefore, hard to detect. Second, FCMs differ in sensitivity and provide data in arbitrary units, which complicates data interpretation. Due to the mentioned challenges, the measured concentration of EVs by flow cytometry is cumbersome to compare between FCMs and institutes. To improve comparability, standardization and development of traceable reference materials to calibrate all aspects of an FCM are needed, as are interlaboratory comparison studies. Within this article, we will provide an overview of the standardization of EV concentration measurements, including the current effort to introduce robust calibration of FCMs, thereby enabling comparable concentration measurements of EVs, which in turn can be used to establish clinically relevant reference ranges of EV concentrations in blood plasma and other body fluids.
KW - calibration
KW - extracellular vesicles
KW - flow cytometry
KW - interlaboratory comparison study
KW - standardization
UR - http://www.scopus.com/inward/record.url?scp=85162203743&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.jtha.2023.04.042
DO - https://doi.org/10.1016/j.jtha.2023.04.042
M3 - Review article
C2 - 37201724
SN - 1538-7933
VL - 21
SP - 2032
EP - 2044
JO - Journal of thrombosis and haemostasis : JTH
JF - Journal of thrombosis and haemostasis : JTH
IS - 8
ER -