Abstract: An extracellular vesicle-containing sample can be processed using a device for isolating one or more subpopulations of the extracellular vesicles. The extracellular vesicle-containing sample is flowed through a flow chamber of the device under an applied fluid pressure, in which the device has one or more inlets and two or more outlets in fluid communication with one another via the flow chamber. The device has one or more filters in the flow chamber between the inlet(s) and at least one of the outlet(s). The extracellular vesicle-containing sample is flowed through the filter(s) in the flow chamber to sort the extracellular vesicles of extracellular vesicle-containing sample by size into two or more subpopulations of the extracellular vesicles. At least one of the subpopulations that has been sorted flows out of a corresponding one of the outlets. Surface marker-based exosome sorting using magnetic beads may be used after the size-based exosome isolation.