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. Author manuscript; available in PMC: 2015 Nov 25.
Published in final edited form as: Science. 2015 Aug 28;349(6251):aab3500. doi: 10.1126/science.aab3500

Fig. 3. Live-cell nonlinear structured illumination microscopy based on patterned photoactivation.

Fig. 3

(A) Single time point from a movie of the evolution of cortical f-actin in a COS-7 cell at 23°C transfected with Skylan-NS-Lifeact, seen at 62-nm resolution (Movie 6, fig. S31, and movie S8). (B) Magnified view from a different cell at 37°C, comparing diffraction-limited TIRF microscopy (top left), TIRF with deconvolution (top right), TIRF-SIM (bottom left), and nonlinear TIRF-SIM with patterned activation (PA NL-SIM, bottom right) (movies S9 and S10). (C) Caveolae in a COS-7 cell at 23°C transfected with Skylan-NS-caveolin, comparing TIRF with deconvolution (top left, 220-nm resolution), TIRF SIM (top right, 97-nm resolution), PA NL-SIM (bottom left, 62-nm resolution), and saturated PA NL-SIM (bottom right, 45-nm resolution). (Insets) A single caveolae pit eventually resolved as a ring by saturated PA NL-SIM (Movie 7, figs. S34 to S37, and movie S13. (D) Diversity of caveolae ring diameters as seen by means of PA NL-SIM. (E) Larger rings that may represent surface-docked vesicles. (F) Clusters of caveolae reminiscent of clathrin plaques. (D) to (F) are from a different cell at 37°C (fig. S33 and movie S12). Scale bars, 3 μm (A); 1 μm (B); 200 nm (C); and 100 nm (D), (E), (F), and (C), inset.