Fig. 5.

(A) Representative SM emission traces of enzymatically generated fluorophores in B. subtilis showing photons detected per 100 ms bin vs. time. For each graph, the inset is a single imaging frame from the specific B. subtilis cell (cell outline in dashed green, SM plotted is in blue box). (B) DL average image of 3000 frames (24 s) of B. subtilis incubated with 1 and corresponding white light image (inset). (C) SR reconstruction (average 17.9 ± 0.3 nm statistical precision) from the cell boxed in blue with both membrane localizations and punctate spots (blue dashed) consisting of many overlapping localizations. (D) Temporal progression of the localizations in the punctate spots, with early localizations colored purple, and later localizations colored white. The lack of temporal progression in the localizations suggests that these punctate spots correspond to the fluorescent cloud generated by two relatively stationary enzymes. (E) SR reconstruction from the green-boxed cell. (F) Temporal progression of localizations in the green dashed box of (E) showing a defined temporal progression with later localizations toward the center of the cell (shown by arrow), suggesting that this punctate spot is a single enzyme diffusing on the time scale of our experiment. Scale bars: 1 μm except 0.5 μm in (D and F). For more details on incubation, imaging, fitting, and display conditions, see the ESI.†