Fig. 6: Two-color live-cell super-resolution microscopy.

a ER (green) and mitochondria (red) of a live Cos7 cell in two-color widefield (left) and SML (right) images. The cell was transfected with UnaG-Sec61β and subsequently stained with MitoTracker Red. The widefield image was taken immediately before super-resolution imaging. b Crosstalk analysis using singly labeled control samples imaged in the two emission channels of green and red split by a dichroic mirror and filtered by a single bandpass filter for each channel. The localizations numbers were normalized by the average count from the proper color channel. c–f Mitochondrial matrix (green) and inner membrane (red) in two-color SML images of a live Cos7 cell transfected with Mito-UnaG and stained by MitoTracker Red. c A far view comparison between widefield (left) and SML (right) images. d Close-up comparison between widefield (left) and SML (right) images of yellow-boxed region in c. e, f Zoom-ins in the cyan- and white-boxed regions in c at 0 min (left) and after 5 min (right) with fusion (white arrows) and fission (yellow arrows) events. Scale bars: 2 µm for a, c; 200 nm for d, f. Error bars: standard deviations (n = 5, from independent SML images).