Fig. 2

Multi-color STORM images for the various compositions of EVs in gram-positive bacteria. A Representative multi-color STORM images of S. aureus without EVs (top) and with EVs (bottom). Red: WGA, protein A, or enterotoxin B. Green: Nile red. B Transverse profiles of localizations corresponding to regions boxed in white in (A). Green bars: Localization frequency measured from the STORM image of Nile red. Red bars: Localization frequency measured from the STORM image of WGA, protein A, or enterotoxin B. C Population ratio of EVs displaying one color (Nile red only) or two colors (Nile red and WGA/ protein A/enterotoxin B) measured from the EVs on the bacterial surface (mean±SD; n=~30). D Comparison of averaged localization density of WGA, protein A, and enterotoxin B of EV-producing S. aureus. E Representative correlative multi-color STORM and SEM images of S. aureus with EV precursors or finalized EVs. F Representative TEM image of S. aureus with EV precursors or finalized EVs. G Representative multi-color STORM images of S. aureus with EVs produced after the explosive cell lysis event. Red: WGA, protein A, or enterotoxin B. Green: Nile red. H Representative TEM image of S. aureus with produced EVs after the explosive cell lysis event. I Population ratio of EVs displaying one color (Nile red only) or two colors (Nile red and WGA/ protein A/enterotoxin B) measured from the STORM images of the released EVs after the explosive cell lysis event (n=~20). White arrows: observed EVs. Scale bars: 1 μm in A, E, and G and 500 nm in F and H