Fig. 5: me4Pi-SMLM live-cell imaging.

a, Super-resolution image of the ER in a live U-2 OS cell recorded over 10 min. Colour bar denotes time. b, 3D super-resolution reconstruction of the ER from images acquired over 10 s. Colour bar denotes depth. c,d, Time-lapse images of ER dynamics from the boxed regions denoted in b. e,f, Lateral and axial intensity profiles along the magenta line denoted in b were fitted with Gaussian functions to estimate the full width at half maximum (FWHM) of the ER tubule. g, FRC analysis of the image shown in b. h, Single-molecule image of the ER in a live U-2 OS cell. Green circles indicate the positions of single molecules. i, Cumulative image of single-molecule trajectories over 30 s. Tracks comprising fewer than ten frames are not displayed. Colour bar denotes time. j, Representative 3D single-molecule trajectories showing different molecular mobility. k, Distribution of localization precision in each dimension. l, Histogram of track duration. m, Histogram of total displacement. n, Histogram of mean speed. o, Histogram of confinement ratio. Representative images are shown from four to five independent experiments.