Figure 1.

Influence of cluster size and density on interpretation of dSTORM experiments. (A) Fluorescence of a single, immobilized Alexa647-labeled secondary antibody in buffer containing 100 mM MEA for 20,000 frames at 20 Hz. Fluorophore blinking seen as upward ticks in fluorescence intensity. (B) Distribution of on-time per blink (t_on), off-time per blink (t_off), number of blinks/antibody, and emission rate measured for 297 individual blink events from 99 individual labeled antibodies. (C) Superresolution images generated from simulations for AQP4 under disperse conditions, in small clusters (30 tetramers) and in large clusters (90 tetramers), at low (300 tetramers/μm²) and high (3000 tetramers/μm²) AQP4 density. (D). Blinking-corrected pair-correlation function, g(r), calculated at 10-nm intervals from the data sets in (C) (solid circles) and fitted to a single exponential (solid line). (E) Cluster size calculated from fitted amplitude and exponential time constant of g(r) as a function of AQP4 density, for small and large clusters (dotted lines). To see this figure in color, go online.