title: Imaging flow cytometry for the characterization of extracellular vesicles.
abstract: Extracellular Vesicles (EVs) are potent bio-activators and inter-cellular communicators that play an important role in both health and disease. It is for this reason there is a strong interest in understanding their composition and origin, with the hope of using them as important biomarkers or therapeutics. Due to their very small size, heterogeneity, and large numbers there has been a need for better tools to measure them in an accurate and high throughput manner. While traditional flow cytometry has been widely used for this purpose, there are inherent problems with this approach, as these instruments have traditionally been developed to measure whole cells, which are orders of magnitude larger and express many more molecules of identifying epitopes. Imaging flow cytometry, as performed with the ImagestreamX MKII, with its combination of increased fluorescence sensitivity, low background, image confirmation ability and powerful data analysis tools, provides a great tool to accurately evaluate EVs. We present here a comprehensive approach in applying this technology to the study of EVs.
authors.affiliation: University of Virginia, School of Medicine, Flow Cytometry Core, 1300 Jefferson Park Avenue, Charlottesville, VA 22908-0734, USA. Electronic address: jl7fj@virginia.edu.
authors.forename: Joanne
authors.initials: J
authors.lastname: Lannigan
authors.affiliation: University of Virginia, Department of Medicine/Nephrology Division, 1300 Jefferson Park Avenue, Charlottesville, VA 22908-0133, USA. Electronic address: ue2u@virginia.edu.
authors.forename: Uta
authors.initials: U
authors.lastname: Erdbruegger
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002450:Cell Communication; D055252:Cell-Derived Microparticles / Q000378:metabolism / Q000648:ultrastructure*; D003713:Dendritic Cells / Q000378:metabolism / Q000648:ultrastructure; D055354:Exosomes / Q000378:metabolism / Q000648:ultrastructure*; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation / Q000379:methods*; D018969:Insulin-Like Growth Factor Binding Proteins / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D013194:Staining and Labeling / Q000379:methods*; D013388:Succinimides / Q000737:chemistry; D014584:User-Computer Interface
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D015415:Biomarkers; D005452:Fluoresceins; D005456:Fluorescent Dyes; D018969:Insulin-Like Growth Factor Binding Proteins; D013388:Succinimides; C102529:insulin-like growth factor binding protein-related protein 1
keywords: Exosomes; Extracellular vesicles; Imagestream; Imaging flow cytometry; Microparticles