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Figure 4. Free flow electrophoresis (FEE) allows preparation of bona fide extracellular vesicles (EVs). FFE fractions considered to contain the majority of EVs were analyzed by nanoparticle tracking analysis (NTA), Western blot (WB), and Transmission electron microscopy (TEM). (A) The results of NTA analyses depicted as particles per mL and mean particle size (131.4 nm) of Fractions 7-10 of the FFE fractioned MSC-CM (for details, see CM1 in Table 1). (B) Anti-CD9 Western blot of the same fractions depicted in (A). (C) TEM images of detectable Fraction 9 components following uranylacetate fixation: (top) 10,000× (1.189 nm/px) magnification (120 kV); and (bottom) 50,000× (0.240 nm/px) magnification.