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

Yohan Kim, Edwin van der Pol, Ali Arafa, Ishwor Thapa, Cameron J. Britton, Jorgena Kosti, Siyang Song, Vidhu B. Joshi, Ree M. Erickson, Hesham Ali, Fabrice Lucien

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Pages (from-to)9781-9795
Number of pages15
Issue number27
StatePublished - May 31 2022

ASJC Scopus subject areas

  • Materials Science(all)


Dive into the research topics of 'Calibration and standardization of extracellular vesicle measurements by flow cytometry for translational prostate cancer research'. Together they form a unique fingerprint.

Cite this