Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry

Leonie de Rond, Edwin van der Pol, Chi M. Hau, Zoltan Varga, Auguste Sturk, Ton G. van Leeuwen, Rienk Nieuwland, Frank A. W. Coumans

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69 Citations (Scopus)


Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. +)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. +EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used
Original languageEnglish
Pages (from-to)680-689
JournalClinical Chemistry
Issue number4
Publication statusPublished - 2018

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