Abstract The importance of cell-derived small particles has recently been recognized in multiple biological processes and is an increasing topic of investigation in biomedical research. Extracellular vesicles (EV) are cell-derived membranous vesicles that include exosomes, microvesicles, and apoptotic bodies. Small particle analysis by flow cytometry can bring much needed discoveries in small particle research, with the capability to detect rare events in significant numbers as well as measure surface protein levels. However, traditional flow cytometers were not capable of resolving them due to their small diameter and variable composition. The aim of this investigation is to determine the capacities of the NovoCyte QuanteonTM flow cytometer to resolve small particles, using both beads and cell-derived EV. We first performed analysis of three commercially available small particle bead mixes ranging in size of 100nm-1300nm and determined the Quanteon is able to resolve beads as small as 100nm. Since beads scatter more light than similar sized membranous particles, it is important to measure biological samples to test the detection limits of the Quanteon. Therefore, HEK293 derived GFP lipoparticles were analyzed for scatter resolution and GFP expression. In addition, plasma was analyzed for purified EV with and without ADP addition, a molecule known to induce platelet activation and EV release. Next, EV from a cancer cell line, SW480, was analyzed for known EV markers CD9, CD81, and AnnexinV. The Quanteon was able to effectively analyze lipoparticles and secreted EV. In conclusion, the NovoCyte Quanteon equipped with new technologies allows better discrimination of small particles and provide increased capacity for research into EV.