Fig. 2: Demonstration of SIMFLUX on DNA-origami nanostructures.
From: Localization microscopy at doubled precision with patterned illumination
a, Full 26-μm wide FOV SIMFLUX image of sparsely distributed nano-rulers with 80-nm spacing. Four independent imaging experiments were done with similar outcome. Scale bar, 5 μm. b,c, Zoom-in on four conventional SMLM (b) and SIMFLUX (c) nano-ruler instances color indicated as boxes in a, both reconstructions are based on the same underlying data. Scale bars, 50 nm. d,e, SMLM (d) and SIMFLUX (e) image of nano-ruler instance of box in a. Scale bars, 50 nm. f,g, Histograms of localizations in d,e projected on the x axis. h,i, Two-dimensional histograms of SMLM (h) and SIMFLUX (i) localizations in the image plane, assembled from 420 segmented binding sites. Scale bars, 15 nm. j,k, Histograms of of SMLM (j) and SIMFLUX (k) localizations projected onto the x direction. l,m, Localization error Δloc and CRLB (mean and s.d.) determined from repeated localizations of long-lasting on-events. Number of localizations per data point are given in the Supplementary Table. n,o, Histogram of nearest neighbor localizations for SMLM (n) and SIMFLUX (o) and bimodal Gaussian fits. p, FRC curves for dataset of a with resolution values R. q,r, SMLM (q) and SIMFLUX (r) images of DNA-origami grids with 40 nm spacing between binding sites. Two independent imaging experiments were done with similar outcome. s,t, SMLM (s) and SIMFLUX (t) images of DNA-origami grids with 20 nm spacing between binding sites. Two independent imaging experiments were done with similar outcomes. Scale bars in q–t, 50 nm.
