Immunophenotyping Extracellular Vesicles using Amnis CellStream

Exosomes are released under normal physiological conditions; however, they are also believed to serve as mediators in the pathogenesis of neurological, vascular, hematological, and autoimmune diseases, as well as cancer. Quantifying and characterizing EVs in a reproducible and reliable manner is difficult due to their small size (exosomes range from 30 nm to 100 nm in diameter). Although EV analysis can be performed using high magnification microscopy, this technique has a very low throughput. Attempts to analyze EVs using traditional PMT-based flow cytometers has been hampered by the limit of detection of such small particles and their low refractive index.To overcome these limitations, we employed the Amnis CellStream flow cytometer, which contains the Amnis Time Delay Integration (TDI) image capturing system. This detection technology allows the CellStream Instrument to combine the advantages of high throughput flow cytometry with high sensitivity to submicron particles.In this study, the CellStream Flow Cytometer was used to immunophenotype EVs derived from red blood cells (RBCs) and platelets.

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