Figure 1.

Examples of conventional methods for EV characterization. a) Different techniques provide information on physical or chemical properties, or a combination of both, and can be further classified based on single particle or ensemble analysis. The fluorescence‐based microfluidic diffusion sizing described in this work is an ensemble multiparametric technique. b) Fluorescence‐based microfluidic diffusion sizing (red box) provides a considerable amount of information on physical and chemical properties of the EV samples while requiring less material and time with respect to the common characterization protocol, i.e., the combination of immunoblotting, nanoparticle tracking analysis, and electron microscopy (black box). Although the microfluidic assay cannot replace the comprehensive characterization with multiple orthogonal techniques, it represents a convenient complementary tool, in particular for fast screening applications and bioprocess development.