Enhancing Analysis of Extracellular Vesicles by Microfluidics

Extracellular vesicles (EVs) play crucial roles in intercellular communication and hold great promise as biomarkers for noninvasive disease diagnosis. Intensive research efforts have been devoted to discovering the EV subpopulations responsible for specific functions or with enhanced effectiveness as disease markers, through extensive EV purification and content analysis. However, their high heterogeneity in size and cargo composition poses significant challenges for reaching such goals. Isolation methods like ultracentrifugation and size-exclusion chromatography, as well as content analysis approaches like polymerase chain reaction and enzyme-linked immunosorbent assay, have made significant contributions to improving our understanding of EV biology. Nonetheless, these methods face limitations in isolation efficiency, EV purity, and detection sensitivity and specificity due to issues like large sample consumption, unsatisfactory purity, and insufficient resolution in EV subtyping. Microfluidic technology presents promising solutions to these challenges, leveraging their intrinsic capabilities in precise flow and external energy field manipulation, sample compartmentalization, and signal enhancement at the micro- and nanoscale. Hence, this review summarizes the recent developments in microfluidics-enabled EV analysis, paying special attention to the unique microfluidic features exploited. Strategies such as viscoelastic and inertial flow, fluid mixing, and external-field-assisted approaches in improving EV purification, as well as compartmentalization and micro/nanostructures for enhancing EV detection, are examined. Furthermore, the current limitations and potential future directions are discussed to inspire advancements in this rapidly developing field.