Figure 6.

Rising opportunities. (a) Improving single-molecule localization via modulating the illumination pattern. (b) In one implementation, the wide-field image is fast scanned between six recording positions, so that six images under different synchronized illumination patterns are concurrently recorded in every frame. (c) Lifetime-resolved SMLM image of two beads labeled by two different dyes, obtained using a confocal setup. (d) Histograms of photon arrival time for two single molecules in (c). Blue lines: exponential fits. (e) Wide-field fluorescence images of single Alexa Fluor 532 molecules, with (top) and without (bottom) gating at 1.6 ns with a Pockels cell. (f) Brightness vs. the estimated lifetime for the molecules numbered in (e), based on the relative intensities in the two views. (g) Wide-field fluorescence images of single C-SNARF-1 molecules in an aluminosilicate film in 580 (top) and 640 (bottom) nm emission channels. (h) Emission ratio image for identified single molecules. (i) SMdM diffusivity map obtained using the non-switchable FP mEmerald. Inset: Distribution of 1-ms single-molecule displacements for a typical region. (j) A convolutional neural network (CNN) receives a raw image of overlapping complex PSFs and outputs a 3D high-resolution volume. (a) is from ref¹⁰⁴. (b) is from ref¹⁰⁵. (c,d) are from ref¹¹². (e,f) are from ref¹¹³. (g,h) are from ref¹³⁵. (i) is from ref⁸⁷. (j) is from ref⁹⁹.