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Fig. 1 | BMC Biology

Fig. 1

From: Visualizing extracellular vesicle biogenesis in gram-positive bacteria using super-resolution microscopy

Fig. 1

Super-resolution images of extracellular vesicles (EVs) from gram-positive bacteria. A 3D STORM images of S. aureus without EVs (top) and with EVs (bottom). S. aureus was labeled with Nile red, WGA, anti-protein A, or anti-enterotoxin B. Inset: Diffraction-limited fluorescence images of the same area. B SEM (left) and TEM (right) images of S. aureus without EVs and with EVs. C The average diameter of EVs measured from STORM (Nile red, WGA, protein A, and enterotoxin B), SEM, and TEM images (mean±SD; n=11–75). D Comparison of the population ratio of S. aureus-secreting EVs observed from STORM images of the sample labeled with Nile red/WGA/anti-protein A/anti-enterotoxin B (mean±SD; n=700–1400). E Correlative STORM and SEM images of EVs labeled with Nile red, WGA, anti-protein A, or anti-enterotoxin B. White arrow: EV observed both from STORM and SEM images. Yellow arrow: EV observed from the Nile red image, which was not shown in the SEM image, implying a membrane vesicle located inside the peptidoglycan layer before the budding process through the cell wall. Blue arrow: EV observed from the SEM image, which was not shown in the STORM image of the cell wall, implying EVs without the peptidoglycan layer. F Fraction of labeled EVs with Nile red, WGA, anti-protein A, or anti-enterotoxin B among the EVs shown in SEM images from correlative STORM and SEM images (n=8–22). G Representative STORM images and the population ratio of Nile red-labeled S. aureus-secreting EVs in the resting state or in the division state. (mean±SD; n=170–540) Scale bars: 1 μm in A, E, and G and 500 nm in B

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