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Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research. / Kim, Yohan; van der Pol, Edwin; Arafa, Ali et al.

In: Nanoscale, Vol. 14, No. 27, 31.05.2022, p. 9781-9795.

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Harvard

Kim, Y, van der Pol, E, Arafa, A, Thapa, I, J. Britton, C, Kosti, J, Song, S, Joshi, VB, M. Erickson, R, Ali, H & Lucien, F 2022, 'Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research', Nanoscale, vol. 14, no. 27, pp. 9781-9795. https://doi.org/10.1039/d2nr01160c

APA

Kim, Y., van der Pol, E., Arafa, A., Thapa, I., J. Britton, C., Kosti, J., Song, S., Joshi, V. B., M. Erickson, R., Ali, H., & Lucien, F. (2022). Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research. Nanoscale, 14(27), 9781-9795. https://doi.org/10.1039/d2nr01160c

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BibTeX

@article{61d44a3e9c614493bac0f5761db248e6,
title = "Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research",
abstract = "Extracellular vesicles (EVs) are microscopic particles released naturally in biofluids by all cell types. Since EVs inherits genomic and proteomic patterns from the cell of origin, they are emerging as promising liquid biomarkers for human diseases. Flow cytometry is a popular method that is able to detect, characterize and determine the concentration of EVs with minimal sample preparation. However, the limited awareness of the scientific community to utilize standardization and calibration methods of flow cytometers is an important roadblock for data reproducibility and inter-laboratory comparison. A significant collaborative effort by the Extracellular Vesicle Flow Cytometry Working Group has led to the development of guidelines and best practices for using flow cytometry and reporting data in a way to improve rigor and reproducibility in EV research. At first look, standardization and calibration of flow cytometry for EV detection may seem burdensome and technically challenging for non-academic laboratories with limited technical training and knowledge in EV flow cytometry. In this study, we build on prior research efforts and provide a systematic approach to evaluate the performance of a high sensitivity flow cytometer (herein Apogee A60-Micro Plus) and fine-tune settings to improve detection sensitivity for EVs. We performed calibration of our flow cytometer to generate data with comparable units (nanometers, MESF). Finally, we applied our optimized protocol to measure the concentrations of prostate-derived EVs in healthy individuals and prostate cancer patients. In conclusion, our proof-of-feasibility study can serve as a scientific and technical framework for other groups motivated in using flow cytometry for EV research.",
author = "Yohan Kim and {van der Pol}, Edwin and Ali Arafa and Ishwor Thapa and {J. Britton}, Cameron and Jorgena Kosti and Siyang Song and Joshi, {Vidhu B.} and {M. Erickson}, Ree and Hesham Ali and Fabrice Lucien",
note = "Funding Information: This work was supported by departmental start-up grant (F. L.) and generous benefactors (F. L.). We also thank generous patients who have participated to the study by providing biospecimens. Publisher Copyright: {\textcopyright} 2022 The Royal Society of Chemistry",
year = "2022",
month = may,
day = "31",
doi = "10.1039/d2nr01160c",
language = "English",
volume = "14",
pages = "9781--9795",
journal = "Nanoscale",
issn = "2040-3364",
publisher = "Royal Society of Chemistry",
number = "27",

}

RIS

TY - JOUR

T1 - Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research

AU - Kim, Yohan

AU - van der Pol, Edwin

AU - Arafa, Ali

AU - Thapa, Ishwor

AU - J. Britton, Cameron

AU - Kosti, Jorgena

AU - Song, Siyang

AU - Joshi, Vidhu B.

AU - M. Erickson, Ree

AU - Ali, Hesham

AU - Lucien, Fabrice

N1 - Funding Information: This work was supported by departmental start-up grant (F. L.) and generous benefactors (F. L.). We also thank generous patients who have participated to the study by providing biospecimens. Publisher Copyright: © 2022 The Royal Society of Chemistry

PY - 2022/5/31

Y1 - 2022/5/31

N2 - Extracellular vesicles (EVs) are microscopic particles released naturally in biofluids by all cell types. Since EVs inherits genomic and proteomic patterns from the cell of origin, they are emerging as promising liquid biomarkers for human diseases. Flow cytometry is a popular method that is able to detect, characterize and determine the concentration of EVs with minimal sample preparation. However, the limited awareness of the scientific community to utilize standardization and calibration methods of flow cytometers is an important roadblock for data reproducibility and inter-laboratory comparison. A significant collaborative effort by the Extracellular Vesicle Flow Cytometry Working Group has led to the development of guidelines and best practices for using flow cytometry and reporting data in a way to improve rigor and reproducibility in EV research. At first look, standardization and calibration of flow cytometry for EV detection may seem burdensome and technically challenging for non-academic laboratories with limited technical training and knowledge in EV flow cytometry. In this study, we build on prior research efforts and provide a systematic approach to evaluate the performance of a high sensitivity flow cytometer (herein Apogee A60-Micro Plus) and fine-tune settings to improve detection sensitivity for EVs. We performed calibration of our flow cytometer to generate data with comparable units (nanometers, MESF). Finally, we applied our optimized protocol to measure the concentrations of prostate-derived EVs in healthy individuals and prostate cancer patients. In conclusion, our proof-of-feasibility study can serve as a scientific and technical framework for other groups motivated in using flow cytometry for EV research.

AB - Extracellular vesicles (EVs) are microscopic particles released naturally in biofluids by all cell types. Since EVs inherits genomic and proteomic patterns from the cell of origin, they are emerging as promising liquid biomarkers for human diseases. Flow cytometry is a popular method that is able to detect, characterize and determine the concentration of EVs with minimal sample preparation. However, the limited awareness of the scientific community to utilize standardization and calibration methods of flow cytometers is an important roadblock for data reproducibility and inter-laboratory comparison. A significant collaborative effort by the Extracellular Vesicle Flow Cytometry Working Group has led to the development of guidelines and best practices for using flow cytometry and reporting data in a way to improve rigor and reproducibility in EV research. At first look, standardization and calibration of flow cytometry for EV detection may seem burdensome and technically challenging for non-academic laboratories with limited technical training and knowledge in EV flow cytometry. In this study, we build on prior research efforts and provide a systematic approach to evaluate the performance of a high sensitivity flow cytometer (herein Apogee A60-Micro Plus) and fine-tune settings to improve detection sensitivity for EVs. We performed calibration of our flow cytometer to generate data with comparable units (nanometers, MESF). Finally, we applied our optimized protocol to measure the concentrations of prostate-derived EVs in healthy individuals and prostate cancer patients. In conclusion, our proof-of-feasibility study can serve as a scientific and technical framework for other groups motivated in using flow cytometry for EV research.

UR - http://www.scopus.com/inward/record.url?scp=85133624653&partnerID=8YFLogxK

U2 - 10.1039/d2nr01160c

DO - 10.1039/d2nr01160c

M3 - Article

C2 - 35770741

VL - 14

SP - 9781

EP - 9795

JO - Nanoscale

JF - Nanoscale

SN - 2040-3364

IS - 27

ER -

ID: 24983585