Small particle flow cytometry using 3 light scatter detectors enhances extracellular vesicle analysis in liquid biopsies, highlighting the potential to segregate EVs by refractive index
Pink, Desmond, Basu, Arghya, Valencia, Juliana, Pham, Diana, and Lewis, J.D.
Extracellular vesicle analysis using “small particle” flow cytometry would be greatly enhanced if data from materials of different refractive index (RI) could be segregated. Likewise, relative sizing of EVs using small particle flow cytometry is confounded by the influence of RI on light scatter. Beads of different composition and refractive index scatter light differently, so that small beads of high RI and large beads of lower RI can have overlapping signals on a two dimension light scatter plot. As particle size decreases, light scatter intensity profiles eventually merge regardless of refractive index. In this project, we aimed to demonstrate graphically, (1) the enhancement of EV flow analysis when using an additional angle of light scatter collection (medium angle of light scatter, MALS) to identify different sample components (e.g. lipids, protein, extracellular vesicles) and (2) the practical reality of sample component overlap at different particle sizes.
Nanostics CSO Desmond Pink will be presenting this work at CYTO 2022, June 3-7, in Philadelphia, US.
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