Extended Data Fig. 7: Three-dimensional localization distribution of single molecules.
From: Molecular-scale axial localization by repetitive optical selective exposure
a, Three-dimensional localization distribution of single molecules with ROSE-Z. The results were based on the microtubule network data in Fig. 2. Molecules appeared for more than 10 frames were selected for analysis, and localizations from 474 clusters were aligned by their center-of-mass to generate the overall 3D presentation of the localization distribution. b-d, Localization distribution of single molecules in the X- (b), Y- (c) and Z- (d) directions with ROSE-Z, respectively. The histograms were fitted to a Gaussian function, yielding standard deviation of 8.4 nm in X, 8.7 nm in Y and 3.4 nm in Z, respectively. e, Three-dimensional localization distribution of single molecules with astigmatic method. Molecules appeared for more than 10 frames were selected for analysis, and localizations from 387 clusters were aligned by their center-of-mass to generate the overall 3D presentation of the localization distribution. f-h, Localization distribution of single molecules in the X- (f), Y- (g) and Z- (h) directions with astigmatism, respectively. The histograms were fit to a Gaussian function, yielding standard deviation of 8.8 nm in X, 8.7 nm in Y and 17.6 nm in Z, respectively.
