Fig. 6: 4Pi-SIM visualized organelle interactions with simultaneous two-color imaging.
From: Elucidating subcellular architecture and dynamics at isotropic 100-nm resolution with 4Pi-SIM
a, Two-color image (maximum intensity projection) of the ER (dCyOFP2s-KDEL, magenta) and mitochondria (oxStayGold-OMP25, green) in a live HeLa cell (Supplementary Video 15). b, Top: magnified x–y views of the boxed region in a at the indicated time points, showing the ER touching a mitochondrial nanotunnel (denoted by the arrowhead). Bottom: x–z cross-sections along the dashed lines. Right: y–z cross-sections at the colored dashed lines drawn in the 116-s x–y view. c,d, Top: magnified x–y views of the boxed regions in a at the indicated time points, showing a mitochondrial fusion (c) and fission (d) event. Arrowheads indicate the fusion and fission sites. Bottom: x–z cross-sections along the dashed lines. e, Two-color image (maximum intensity projection) of the ER (dCyOFP2s-KDEL, magenta) and microtubules (oxStayGold-Ensconsin, green) in a live COS-7 cell (Supplementary Video 16). f, The y–z cross-section of the dashed line in e. g,h, Top: magnified x–y views of the boxed regions in e at the indicated time points. Arrowheads indicate the contact sites (g) and the coordinated motion of the ER and microtubules (h). Bottom: x–z cross-sections along the dashed lines. Scale bars: 2 μm (a,e) and 1 μm (b–d,f–h). The thickness of all cross-section slices is 30 nm. Representative results are shown from two to four independent experiments.
