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
Related in: MedlinePlus
Mentions: Next, we investigated the influence of Anx5-F* concentration on the detection of Anx5-positive EVs. For both FS and FL triggering, the number of Anx5-positive EVs detected was found to increase with increasing Anx5-F* concentration, until a plateau value was reached (Fig.2). In the conditions used here, namely 10 ×-diluted PFP, the plateau was reached at 0.8 nm Anx5-F*. Most experiments reported in this paper were performed with 2.8 nm Anx5-F*, therefore in saturating conditions.
Affiliation: Molecular Imaging and NanoBioTechnology, UMR-5248-CBMN CNRS-University of Bordeaux-IPB, Pessac, France.