Application of Flow Cytometry in the Studies of Microparticles

The Methods of Choice for Extracellular Vesicles (EVs) Characterization

“…On the other hand, side-scattered light (15–150°) is often collected at the 90° angle and provides information about smaller particles or granularity of internal structures. Thus, the side scatter (SSC) has been proposed as a trigger channel [ 36 ], as most cytometers show a better detection sensitivity using SSC as a trigger rather than FSC for the analysis of small particles [ 36 , 37 ]. Using SSC in this manner enables reproducing the sensitivity level of 190 nm latex particles [ 36 ], however, it can still lead to problems associated with EVs resolution above the background noise [ 38 ].…”

“…As a result, the vesicles population studied can be quantified and/or classified according to the level of antigen expression ( Figure 2 B) [ 43 ]. Flow cytometry can also be used to enumerate EVs by adding, as an internal standard, a known number of fluorescent latex beads (Flow-Count Fluorospheres, Beckmann-Coulter, Brea, CA, USA) or using tubes containing already predefined bead numbers (TruCount tubes, BD Biosciences, San Jose, CA, USA) [ 37 ]. For smaller particles, such as exosomes (<100 nm in diameter) some other approaches can be introduced to allow their analysis by flow cytometry.…”