Analysis of extracellular vesicles by flow cytometry – basics, limitations and prospects

Abstract

Flow cytometry is a well-established technique that classically is used to detect cells and quantify related parameters on the cellular surface, e. g. the expression of surface protein markers. Within the last few years, there also have been considerable advances of using flow cytometry to detect extracellular vesicles (EVs) in the size-range of exosomes and quantify their concentration and surface marker expression in EV-containing samples such as cell culture supernatant or biological fluids. Of note, such analyses of EVs within a size range well beyond <300 nm are still challenging and require a suitable instrumentation and a certain level of expertise to make sure essential controls are included and to ensure that resulting datasets can be interpreted appropriately. After giving a brief, basic introduction into flow cytometry which is essential especially for non-experienced readers in order to understand submicron particle flow cytometry data, we will give a concise overview about current possibilities and methods available in the field.