Extracellular vesicles (EVs) have gained significant interest as potential bio-markers and bio-activators in health and disease. These submicron vesicles are believed to transfer proteins, lipids and nucleic acids, thus facilitating communication between cells. The detection of this wide range of EVs is challenged by their small and heterogeneous size range, high concentration, low refractive index, and heterogeneity in composition and morphology. This protocol uses imaging flow cytometry (IFC) to detect EVs as it combines high fluorescence sensitivity, low background, image confirmation ability and powerful data analysis tools. As EVs carry markers of their parent cells, IFC can also be used to identify EV origin via targeted and high-throughput phenotyping.
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