Single Extracellular Vesicle Transmembrane Protein Characterization by Nano-Flow Cytometry

Single particle characterization has become increasingly relevant for research into extracellular vesicles, progressing from bulk analysis techniques and first-generation to comprehensive multi-parameter measurements such as nano-flow cytometry (nFCM). nFCM is a form of flow that utilizes instrumentation specifically designed nano-particle analysis, allowing thousands EVs be characterized per minute both with without the use staining techniques. High resolution side scatter (SS) detection allows size concentration determined all biological particles larger than 45 nm, while simultaneous fluorescence (FL) identifies presence labeled markers targets interest. Labeled subpopulations can then described in quantitative units particles/mL or percentage total identified by scatter. Here, derived conditioned cell culture media (CCM) are lipid dye, identify membrane, antibodies specific CD9, CD63, CD81 common EV markers. Measurements comparison material, standard silica nanospheres, well sample material analyzed 1-minute analysis. The software used measure distribution profile particles, independent labeling, before determining positive each labels. Simultaneous SS FL utilized flexibly many different sources labeling targets, external internal, describing samples manner.