A simple flow cytometry method improves the detection of phosphatidylserine-exposing extracellular vesicles.
Bottom Line: We show that about 50× more Anx5-positive EVs are detected by FCM when detection is triggered on fluorescence as compared with light scattering.By fluorescence triggering, concentrations of 22 000-30 000 Anx5-positive EVs per μL PFP were determined, using two different flow cytometers.Results from EM suggest that EVs down to 100-150 nm diameter are detected by fluorescence triggering.
Affiliation: Molecular Imaging and NanoBioTechnology, UMR-5248-CBMN CNRS-University of Bordeaux-IPB, Pessac, France.Show MeSH
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Mentions: Finally, we attempted to phenotype the Anx5-positive EVs detected by FL triggering. We focused on the populations of EVs derived from platelets and erythrocytes, which were identified by labeling with anti-CD41-mAb-PE and anti-CD235a-mAb-PE, respectively. As shown in Fig.6, when PFP was double labeled with Anx5-Cy5 and either anti-CD41-PE (Fig.6B) or anti-CD235a-PE (Fig.6C), a population of events positive for both markers was observed in the FL6 vs. FL2 color dot plots. We found that, for this PFP sample, 23% and 8% of the Anx5-positive EVs detected by FL triggering derived from platelets and erythrocytes, respectively.
Affiliation: Molecular Imaging and NanoBioTechnology, UMR-5248-CBMN CNRS-University of Bordeaux-IPB, Pessac, France.