%0 Journal Article
%T High-fidelity detection and sorting of nanoscale vesicles in viral disease and cancer
%A Aizea Morales-Kastresana
%A Alan Dean
%A Aliaks
%A Anu Puri
%A Aparna H. Kesarwala
%A Ariel Rosner
%A Bryant Yung
%A Carley D. Ross
%A David R. Parks
%A Edward J. Felton
%A Gordon J. Freeman
%A Hisataka Kobayashi
%A Ionita Ghiran
%A James C. S. Wood
%A Jane B. Trepel
%A Jay A. Berzofsky
%A Jennifer C. Jones
%A John Tigges
%A Jonathan Van Dyke
%A Joshua A. Welsh
%A Katherine McKinnon
%A Lise Pasquet
%A Marjorie Robert-Guroff
%A Marty Bigos
%A Masaki Terabe
%A Peter Choyke
%A Peter Guion
%A Stephen P. Perfetto
%A Susan J. Knox
%A Thomas A. Musich
%A Thorsten Demberg
%A Vasilis Toxavidis
%A Veena Kapoor
%A W. Roy Overton
%A William Telford
%A Xiaoyuan Chen
%A er Kachynski
%J Journal of Extracellular Vesicles
%D 2019
%R https://doi.org/10.1080/20013078.2019.1597603
%X ABSTRACT Biological nanoparticles, including viruses and extracellular vesicles (EVs), are of interest to many fields of medicine as biomarkers and mediators of or treatments for disease. However, exosomes and small viruses fall below the detection limits of conventional flow cytometers due to the overlap of particle-associated scattered light signals with the detection of background instrument noise from diffusely scattered light. To identify, sort, and study distinct subsets of EVs and other nanoparticles, as individual particles, we developed nanoscale Fluorescence Analysis and Cytometric Sorting (nanoFACS) methods to maximise information and material that can be obtained with high speed, high resolution flow cytometers. This nanoFACS method requires analysis of the instrument background noise (herein defined as the ˇ°reference noiseˇ±). With these methods, we demonstrate detection of tumour cell-derived EVs with specific tumour antigens using both fluorescence and scattered light parameters. We further validated the performance of nanoFACS by sorting two distinct HIV strains to >95% purity and confirmed the viability (infectivity) and molecular specificity (specific cell tropism) of biological nanomaterials sorted with nanoFACS. This nanoFACS method provides a unique way to analyse and sort functional EV- and viral-subsets with preservation of vesicular structure, surface protein specificity and RNA cargo activity
%U https://www.tandfonline.com/doi/full/10.1080/20013078.2019.1597603