Exploring exosome profiling via CytoFLEX Nano flow cytometer: Approaches and applications

Abstract Exosomes, a subtype of extracellular vesicles, have garnered significant attention as biomarkers and therapeutic targets. Recent advances in flow cytometry have enhanced the characterization of individual particles and improved the analysis of extracellular vesicle populations. This study focused on the CytoFLEX Nano flow cytometer, designed for small particle detection. We assessed the particle size distribution and marker protein profiles of exosome samples, comparing those from different sources and isolated using various methods to examine the stability and heterogeneity of exosome markers. The results revealed that the CytoFLEX Nano effectively characterized exosome samples, differentiated those from various sources, and emphasized the considerable influence of different separation methods on exosome labeling and enrichment efficiency. Moreover, the CytoFLEX Nano exhibited higher sensitivity and better labeling efficiency than the CytoFLEX for detecting smaller particles. In conclusion, these findings help optimize instrument selection and experimental design, thereby supporting their broad application in fundamental research, disease monitoring, and clinical diagnostics.