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. Author manuscript; available in PMC: 2018 Aug 28.
Published in final edited form as: Chem Rev. 2018 Jan 31;118(4):1917–1950. doi: 10.1021/acs.chemrev.7b00534

Figure 7. Single EV analysis (SEA).

Figure 7

(a) EVs are biotinylated and captured on a flat surface coated with neutravidin (Av). EVs are then stained with fluorescent antibodies and imaged. Subsequently, fluorophores are quenched and the staining process is repeated for a different set of markers. (b) Example SEA image. EVs from Gli36-WT cell line were biotinylated and captured. Individual EV were detected through staining with fluorescent streptavidin StAv (top left). For molecular profiling, EVs were stained for pan-EV markers (tetraspanins; CD9, CD63, CD81) as well as tumor markers (EGFR, EGFRvIII, IDH1, IDH1R132). Spots with circles indicate individual EVs. (c) 2-dimensional tSNE mapping of individual EVs analyzed for protein markers. The original map was redrawn for EVs from a single cell line. Data from other cell lines are shaded light gray. EVs from Gli36-WT and Gli36-IDH1R132 cells lines clustered similarly, whereas EVs from Gli36-EGFRvIII cells showed distinct clusters. Reprinted with permission from Ref 82. Copyright 2017 American Chemical Society.