Microfluidics for separation, detection, and engineering of extracellular vesicles

This review presents a clear and engaging exploration of how microfluidic technologies are reshaping the field of extracellular vesicle (EV) research, bridging technical innovation with clinical relevance. It aims to connect fundamental microfluidic principles, such as laminar flow and nanoscale manipulation, with their practical applications in enhancing the separation, detection, and engineering of EVs. The goal is to show not only how these systems work but also why they matter for advancing diagnostics, therapeutics, and precision medicine. The review describes a range of isolation strategies, including immunoaffinity capture, size-based nanofiltration, acoustic separation, and dielectrophoresis, illustrated by prominent examples like ExoChip, Exodisc, and Nano-IMEX. These devices achieve higher recovery rates, purity, and throughput compared with traditional ultracentrifugation, all while requiring smaller sample volumes and less processing time. The review also highlights emerging detection tools that integrate microfluidics with advanced biosensing modalities, including electrochemical and impedance-based platforms as well as optical techniques such as surface plasmon resonance and surface-enhanced Raman scattering. Together, these approaches enable sensitive, label-free, and real-time analysis of EVs at the single-vesicle level, providing precise electrical and molecular signatures. Furthermore, the review evaluates microfluidic approaches for loading therapeutic cargo into EVs as drug delivery systems, highlighting their superior precision, efficiency, and scalability over conventional methods. It also describes microfluidic strategies for EV surface modification and integrated bioreactor platforms that combine cell culture, EV production, capture, loading, functionalization, and controlled release within closed, automated systems. It is worth noting that the review also assesses challenges in device scaling and discusses the future opportunities ranging from AI-driven signal interpretation to standardized, scalable fabrication that could accelerate real-world adoption. Ultimately, microfluidic technologies are redefining medicine by enabling faster, more precise diagnostics and targeted therapies.