Extended Data Fig. 2: Characterizing properties of the pre-RL deconvolution.

(a) A representative image of a single molecule with a cosine intensity fluctuation convolved by a 220 nm Gaussian PSF before adding Poisson noise only. (b) Normalized intensity profiles of ground-truth (black), raw (blue), and RL deconvolution (red) for the temporal response indicated by the black point in (a). (c) A representative image of two blinking molecules convolved by a 220 nm Gaussian PSF before adding Poisson noise, ~2% Gaussian noise, and background noise. (d) Intensity profiles of ground-truth (black), raw (blue), and RL deconvolution (red, divided by 3.5804) for the temporal response indicated by the black point (left) and green point (right) in (c). (e) A single molecule convolved by a 155 nm Gaussian PSF as ground-truth (‘GT’; left), and convolved by a 220 nm Gaussian PSF before adding Poisson noise (‘Raw’; middle) followed by RL deconvolution (‘RL’; right). (f) Intensity profiles of ground-truth (black), raw (blue), and RL deconvolution (red). Black arrows indicate the pixel-level fluctuations in the raw capture (blue). (g) Representative images of two molecules with opposite cosine intensity fluctuations convolved by a 220 nm Gaussian PSF before adding Poisson noise, ~2% Gaussian noise, and background noise. From left to right: Ground-truth molecules in xy; Colored rapture in xy; Colored raw capture in xt; Uncolored raw capture in xy. (h) Normalized intensity profiles of ground-truth (black), raw (blue), and RL deconvolution (red) for the temporal response indicated by the magenta point (dashed lines) and green point (solid lines) in (g, 1^st column). Details in Supplementary Note 1. Experiments were repeated ten times independently with similar results. Scale bars: (a, e, g) 100 nm; (c) 500 nm.