Advances in multivariate analysis of extracellular vesicles using visible light-based analytical methods

Extracellular vesicles have emerged as promising candidates for diagnostic and therapeutic applications, however their inherent heterogeneity presents significant challenges for their characterization and implementation. A number of visible light-based analytical techniques have emerged to examine the physicochemical properties of extracellular vesicles in ensembles and as individual entities. We discuss how these methods can be utilized to analyze multiple parameters simultaneously, providing a more comprehensive understanding of the distribution of extracellular vesicle properties. These techniques offer insights into the interconnected forms of heterogeneity, including the interplay between size, biomolecular composition, and interactions. This review surveys the current landscape of multivariate heterogeneity analysis, highlighting its potential for advancing extracellular vesicle-based therapies.