Extracellular vesicles (EVs) are complex lipid membrane
vehicles
with variable expressions of molecular cargo, composed of diverse
subpopulations that participate in the intercellular signaling of
biological responses in disease. EV-based liquid biopsies demonstrate
invaluable clinical potential for overhauling current practices of
disease management. Yet, EV heterogeneity is a major needle-in-a-haystack
challenge to translate their use into clinical practice. In this review,
existing digital assays will be discussed to analyze EVs at a single
vesicle resolution, and future opportunities to optimize the throughput,
multiplexing, and sensitivity of current digital EV assays will be
highlighted. Furthermore, this review will outline the challenges
and opportunities that impact the clinical translation of single EV
technologies for disease diagnostics and treatment monitoring.