Quantitative flow cytometry enables end-to-end optimization of cross-platform extracellular vesicle studies.

Autor:	Cook S; Laboratory of Pathology, Translational Nanobiology Section, Centre for Cancer Research, National Institute of Health, National Institutes of Health, Bethesda, MD, USA., Tang VA; Faculty of Medicine, Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Flow Cytometry and Virometry Core Facility, Ottawa, ON K1H 8M5, Canada., Lannigan J; Flow Cytometry Support Services, Alexandria, VA, USA., Jones JC; Laboratory of Pathology, Translational Nanobiology Section, Centre for Cancer Research, National Institute of Health, National Institutes of Health, Bethesda, MD, USA., Welsh JA; Laboratory of Pathology, Translational Nanobiology Section, Centre for Cancer Research, National Institute of Health, National Institutes of Health, Bethesda, MD, USA. Electronic address: joadwe@outlook.com.
Jazyk:	angličtina
Zdroj:	Cell reports methods [Cell Rep Methods] 2023 Dec 18; Vol. 3 (12), pp. 100664.
DOI:	10.1016/j.crmeth.2023.100664
Abstrakt:	Flow cytometry (FCM) is a common method for characterizing extracellular particles (EPs), including viruses and extracellular vesicles (EVs). Frameworks such as MIFlowCyt-EV exist to provide reporting guidelines for metadata, controls, and data reporting. However, tools to optimize FCM for EP analysis in a systematic and quantitative way are lacking. Here, we demonstrate a cohesive set of methods and software tools that optimize FCM settings and facilitate cross-platform comparisons for EP studies. We introduce an automated small-particle optimization (SPOT) pipeline to optimize FCM fluorescence and light scatter detector settings for EP analysis and leverage quantitative FCM (qFCM) as a tool to further enable FCM optimization of fluorophore panel selection, laser power, pulse statistics, and window extensions. Finally, we demonstrate the value of qFCM to facilitate standardized cross-platform comparisons, irrespective of instrument configuration, settings, and sensitivity, in a cross-platform standardization study utilizing a commercially available EV reference material. Competing Interests: Declaration of interests J.A.W. and J.C.J. are inventors on NCI patents and patent applications related to EV analysis. J.L. holds a financial interest in Cytek Biosciences. (Copyright © 2023. Published by Elsevier Inc.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu