Precise analysis of single small extracellular vesicles using flow cytometry

Methods that enable specific and sensitive quantification of small extracellular vesicles (sEVs) using flow cytometry are still under development. Here, we compare four preparation methods for flow cytometry, including ultracentrifugation, density gradient centrifugation, size exclusion chromatography (SEC), and the TIM4-affinity method by analyzing the specificity for antibody staining of sEVs. Conclusion: SEC or the TIM4-affinity method could separate sEVs from antibodies.