DLS and FCM analysis of IC and MP samples. (A) DLS analysis of ICs and MPs, showing both samples are of overlapping size. (B) FCM analysis of ICs, MP, and MPs spiked with ICs. It can be seen the ICs and MPs are distinguished by light scatter alone. (C) FCM analysis of antibody (Ab) aggregate, where Ab aggregate overlaps with the IC population but does not create microvesicle-mimicking signals. For FCM analysis, a Coulter Epics XL instrument was used for acquisition, triggering of side scatter and using a Nano Fluorescent Particle Size Standard Kit; NFPPS-52-4K (Spherotech) for instrument calibration (data not shown). For DLS, a Coulter N4 plus instrument was used, acquiring the data at 90° angle, experiments carried out at 22°C.