Dear Manash!

Please You look at this article:

High sensitivity detection of extracellular vesicles immune-captured from urine by conventional flow cytometry

https://www.nature.com/articles/s41598-019-38516-8

Figure 3

Figure 4

Antibody-coated magnetic beads were obtained from the Exostep™ kit (Immunostep, Ref: ExoS-25-U) or prepared by incubating magnetic fluorescent beads either from Luminex (MagPlex Microsphere, Ref: MC10012) or Bangs (QuantumPlex M SP Carboxil, Ref: 251A) with purified anti-CD63 (Clone TEA3/18), purified anti-CD9 (Clone VJ1/20) or anti-EpCAM (Clone VU-1D9) antibodies from Immunostep, S.L. Magnetic fluorescent beads were coated with high density carboxyl functional groups on the surface, these groups were used to covalently conjugate antibodies through their primary amines by two-step EDC/NHS protocol producing a stable amide bond.

EV flow cytometry

Detection and Quantification of Extracellular Vesicles via FACS: Membrane Labeling Matters!

file://b40.med.local/profiles/RDS_folders/chernovan/Downloads/ijms-21-00291-v2%20(1).pdf

Figure 1 (D) Gating strategy for the identification of intact hsEVs by high-resolution flow cytometry

Hello Manash

I recommend thoroughly filtering sheath fluid, PBS and other every other FACS related fluid with 0.2 micron filter. You will be surprised to see how much difference it makes in reducing noise. Using bleach after each exosome run is also recommended.

Best

thanks for the answer. I have started using the same and am getting far better results. U r absolutely right