Detection and Sorting of Extracellular Vesicles and Viruses Using nanoFACS | Zendy

MoralesKastresana Aizea | Zendy; Welsh Joshua A. | Zendy; Jones Jennifer C. | Zendy

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Detection and Sorting of Extracellular Vesicles and Viruses Using nanoFACS

Author(s) -

MoralesKastresana Aizea,

Welsh Joshua A.,

Jones Jennifer C.

Publication year - 2020

Publication title -

current protocols in cytometry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.718

H-Index - 26

eISSN - 1934-9300

pISSN - 1934-9297

DOI - 10.1002/cpcy.81

Subject(s) - extracellular vesicles , flow cytometry , protocol (science) , staining , cd63 , microvesicles , cytometry , microbiology and biotechnology , nanotechnology , biology , computational biology , biophysics , chemistry , materials science , biochemistry , pathology , medicine , microrna , genetics , alternative medicine , gene

Extracellular vesicles (EVs) are sub‐micron‐sized membranous spheres secreted by cells. EVs play a functional role as intercellular communicators and are associated with a number of diseases. Research into EVs is an area of growing interest due their many potential uses as therapeutic agents, as diagnostic and theranostic biomarkers, and as regulators of cellular biology. Flow cytometry is a popular method for enumerating and phenotyping EVs, even though the majority of EVs are below the detection sensitivity of most commercially available flow cytometers. Here, we present optimized protocols for EV labeling that increase the signal‐to‐noise ratio of EVs by removing residual antibody. Protocols for alignment of high‐resolution jet‐in‐air flow cytometers are also provided. Published 2020. U.S. Government. Basic Protocol 1 : Bulk EV staining with CFSE protein binding dye Basic Protocol 2 : Antigen‐specific staining of EV markers with fluorochrome‐conjugated antibodies Basic Protocol 3 : Astrios EQ instrument setup and sample acquisition Basic Protocol 4 : Counting particles and EVs on Astrios EQ with spike‐in reference beads

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