Enumeration of extracellular vesicles by a new improved flow cytometric method is comparable to fluorescence mode nanoparticle tracking analysis

Nanomedicine : Nanotechnology, Biology, and Medicine
Leonardo PasalicVivien M Y Chen

Abstract

Extracellular vesicles (EVs) play a role in a variety of physiological and pathological processes. However, use of EVs as biomarkers has been hampered by limitations of current detection and enumeration methods. We compared fluorescence-threshold flow cytometry (FT-FC) to nanoparticle tracking analysis (NTA) for enumeration of cell culture-derived EVs. FT-FC and NTA utilising fluorescence mode (F-NTA) enumerated similar numbers of EVs stained with a membrane dye PKH67. Both methods were sufficiently sensitive to detect cell-derived EVs above the background of culture medium. Light scatter NTA (LS-NTA) detected 10-100× more particles than either fluorescence-based method but demonstrated poor specificity. F-NTA appeared to have better sensitivity for <100nm vesicles, however, the FT-FC method combined direct enumeration of EVs with high sensitivity and specificity in the >100nm range. Due to wider availability and higher degree of automation and standardisation, FT-FC is a reasonable surrogate to F-NTA for quantification of EVs. Extracellular vesicles are small particles, which can act as tools for intercellular communication. One recent area of interest in EVs is their potentials as biomarkers. In this article, the authors inve...Continue Reading

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Citations

Jun 11, 2016·Cell Communication and Signaling : CCS·Giulia CesiStephanie Kreis
Aug 17, 2016·International Journal of Molecular Sciences·Dragos CretoiuSanda M Cretoiu
Apr 5, 2018·Clinical Hemorheology and Microcirculation·Marc RomanaNigel S Key
Sep 16, 2017·Frontiers in Cell and Developmental Biology·Joshua A WelshNicola A Englyst
Nov 2, 2018·International Journal of Molecular Sciences·Abdelnaby KhalyfaDavid Gozal
Aug 10, 2020·Scientific Reports·Shivani SharmaJames Gimzewski
Nov 28, 2020·International Journal of Laboratory Hematology·Panjaree SiwaponananKovit Pattanapanyasat
Nov 25, 2018·Nanomedicine : Nanotechnology, Biology, and Medicine·Joseph D BazzillJames J Moon
Jul 1, 2021·Journal of Extracellular Vesicles·Zach TroyerJohn C Tilton

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