Extended Data Fig. 2: Comparison of pan-ExM-t immunolabeling with simulated conventional (non-expanded) microscopy.
From: All-optical visualization of specific molecules in the ultrastructural context of brain tissue
pan-ExM-t images were computationally blurred to simulate imaging of equivalent non-expanded samples as described in the Methods Section under the heading Simulations of non-expanded sample images. Contrast was enhanced using ImageJ’s “Enhance Contrast” function (0.35% saturation), and color channels were rendered inverted (pan) and green (Thy1-eGFP) for visualization. Due to the high effective resolution and optical sectioning resulting from 20x expansion, antibody labeling in pan-ExM samples appears much sparser than in standard, non-expanded samples. The simulations illustrate how these samples would appear without expansion, yielding labeling that appears smoother and denser due to the inferior resolution. a-l, anti-GFP labeling (Thy-1 GFP). a-f, pan-ExM-t images. a, c, d, pan channel overlaid with GFP channel (green). b, e, f, GFP channel only. g-l, simulated non-expanded anti-GFP images. g,i,j, pan channel overlaid with GFP channel (cyan). h, k, l, GFP channel only. m-x, anti-PSD-95 labeling. m-r, pan-ExM-t images. m, o, p, pan channel overlaid with PSD-95 channel (cyan). n, q, r, PSD-95 channel only. s-x, simulated non-expanded anti-PSD-95 images. s, u, v, pan channel overlaid with PSD-95 channel (cyan). t, w, x, PSD-95 channel only. All scale bars: 10 μm, not corrected for expansion factor.
