Abstract

Digital Flow Cytometry for the High-Throughput Multiplexed Analysis of Single Extracellular Vesicles and Particles

Daniel Chiu, Professor, University of Washington

Extracellular vesicles and particles (EVPs) play a central role in liquid biopsy, intercellular communication, and disease transmission and progression, and are emerging therapeutic tools. To better understand the biology of EVPs and fully unlock their diagnostic and therapeutic potential, it is critical to access quantitative information regarding their concentration, size, and biological heterogeneities. To meet this need, we have developed a single-molecule sensitive flow platform, which uses a high-throughput 12-channel flow analyzer that detects each and every fluorescent molecule flowing through a microfluidic channel, and enables multiparameter characterization of EVPs, including single-EVP phenotyping, sizing, and the absolute quantitation of EVP concentrations and biomarker copy numbers.  This new flow technology should have a broad range of applications, from analysis of single EVPs such as exosomes or RNA-binding proteins to characterization of therapeutic lipid nanoparticles, viruses, and proteins; it also provides absolute quantitation of non-EVP samples such as dyes, beads, and Ab-dye conjugates.