Supplementary Figure 10: Instant TIRF-SIM resolves Ras microdomains better than diffraction-limited TIRF.
From: Single-shot super-resolution total internal reflection fluorescence microscopy
a) Image taken of GFP-HRas in U2OS cell in diffraction-limited TIRF, i.e. with pinholes and emission microlenses removed. Insets correspond to higher magnification views of blue and red rectangular regions. b) Image taken in the same cell as in (a) but with instant TIRF-SIM. c) Selected image frames taken of GFP-HRas (green) and pDsRed2-ER (magenta) in U2OS cells from instant TIRF-SIM acquisition. d) The same images as (c) but blurred to simulate diffraction-limited images. Arrows highlight two microclusters resolved in instant TIRF-SIM but not in TIRF. See also Fig. 2i, Supplementary Video 6. Scale bars: a) and b): 5 μm; insets, 1 μm; c) and d): 500 nm. Experiments were performed at least 4 times with similar results; representative data are shown.
