A practical approach to extracellular vesicle characterization among similar biological samples

We found that the EVs differ in size and light refractive properties which were demonstrated through differences in their light intensity values. The asymmetrical nature of these biological vesicles produces slightly different concentration and size estimates in repeated analyses, but consistent reproducible data measurements can be obtained when several parameters are taken into consideration. In this study, we determined the effects of replicate number, capture time, flow rate and sample dilutions on EV quantification estimates. Individually, each of these factors influences the accuracy and reproducibility of sample readings. These findings have important implications for direct comparisons of samples to assess EV production or depletion. Here we present a practical approach for establishing an EV characterization protocol for anyone wanting to compare EV content among similar biological samples.