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. Author manuscript; available in PMC: 2017 Jan 4.
Published in final edited form as: Nat Biotechnol. 2016 Jul 4;34(9):987–992. doi: 10.1038/nbt.3625

Figure 2.

Figure 2

Validation of proExM in different mammalian tissue types. (a–d) Low magnification, wide-field images of pre-expansion (top) and post-expansion (bottom) samples of Thy1-YFP mouse brain (a) and vimentin-immunostained mouse pancreas (b), spleen (c), and lung (d). (e) Composite fluorescence image of Tom20 in Thy1-YFP mouse brain imaged with super-resolution structured illumination microscopy (SR-SIM) (green) and proExM (purple) with conventional confocal microscopy with distortion vector field overlaid (white arrows). (f) Pre-expansion SR-SIM image showing boxed region in (a). (g) Post-expansion confocal image of (f). (h) RMS length measurement error as a function of measurement length for proExM vs SR-SIM pre-expansion for Tom20 staining in Thy1-YFP mouse brain (blue line, mean; shaded area, standard deviation; n = 3 mouse brain cortex samples). (i) High magnification, wide-field fluorescence composite image of vimentin in mouse pancreas before (green) and after (purple) expansion with distortion vector field overlaid (white arrows, see methods). (j) Pre-expansion wide-field image showing boxed region in (i). (k) Post-expansion image of (j). (l) Root mean square (RMS) length measurement error as a function of measurement length for proExM vs widefield pre-expansion images for the different tissue types in (b–d) (blue line, mean; shaded area, standard deviation; n = 3 samples from pancreas, spleen, and lung). (m) Composite fluorescence image of vimentin in mouse pancreas imaged with super-resolution structured illumination microscopy (SR-SIM) (green) and proExM (purple) with conventional confocal microscopy with distortion vector field overlaid (white arrows). (n) Pre-expansion SR-SIM image showing boxed region in (m). (o) Post-expansion confocal image of (n). (p) RMS length measurement error as a function of measurement length for proExM vs SR-SIM pre-expansion for vimentin staining in pancreas (blue line, mean; shaded area, standard deviation; n = 4 fields of view from 2 samples). Scale bars: (a) top 200 μm, bottom 200 μm (physical size post-expansion, 800 μm), (b–d) top 500 μm, bottom 500 μm (2.21 mm, 2.06 mm, 2.04 mm, respectively), (e, f) 10 μm, (g) 10 μm (40 μm), (i) 10 μm, (j) 5 μm, (k) 5 μm (20.4 μm), (m) 5 μm, (n) 5 μm, (o) 5 μm (20.65 μm).