| Literature DB >> 35467279 |
Najat Hajji1,2, Chi M Hau1,2, Rienk Nieuwland1,2, Edwin van der Pol3,4,5.
Abstract
Extracellular vesicles (EVs) are lipid membrane enclosed particles that are released from cells into body fluids, such as blood. EVs offer potential new biomarkers of diseases, because the cellular origin, composition, concentration, and function of EVs change in health and disease. The concentration of EVs from specific cell types in blood can be determined with flow cytometry. A flow cytometer measures fluorescence and light scattering signals from single EVs, but only if these signals are sufficiently bright to be detected. Measured concentrations of EVs are therefore only reproducible and comparable if the detection ranges are known and reported in standard units, such as molecules of equivalent soluble fluorophore (MESF) for fluorescence signals and the diameter in nm for scatter signals. The goal of this protocol is to discuss all steps needed to derive the concentration of cell-type specific EVs within a known diameter range and fluorescence range. More specifically, this protocol describes how to determine the concentration of CD61+ (Integrin beta-3, platelet marker), CD235a+ (Glycophorin A, erythrocyte marker), and CD45+ (leukocyte common antigen) EVs in human blood plasma with an Apogee A60-Micro flow cytometer using scatter-based triggering. The principles behind this protocol could lay a firm basis for the design of a protocol suitable for other flow cytometers and body fluids.Entities:
Keywords: Calibration; Extracellular vesicles; Flow cytometry; Fluorescent antibody labeling; Human blood plasma; Number concentration; Standardization
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Year: 2022 PMID： 35467279 DOI： 10.1007/978-1-0716-2341-1_5
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745