Fig. 1
ADAPT-3D tissue processing and refractive index matching render tissues optically transparent without shrinkage. (A) Comparison of fluorescent intensities of CD11c-eYFP follicles in Peyer’s patches of mouse ileum following different fixative conditions and captured by stereomicroscopy. Indicated are averages ± 1 standard error of mean (n = 4–5 follicles per condition, two-way ANOVA followed by Tukey test, *p-value ≤ 0.05, **p-value ≤ 0.01), average). (B) Luminal side of fixed 600 μm thick mouse colons decolorized overnight (left) or left unprocessed (right) shown before (top) and after (bottom) a 60-minute incubation in ADAPT:RI. (C) Full thickness [1.5–2 mm] cross sections of fixed human colon untreated (top) or incubated in ADAPT:RI for 10 min (bottom) without decolorization or delipidation. (D) Fixed mouse and piglet colon incubated for 60 min in ADAPT:DC, 60 min in ADAPT:PDL (left) and then incubated in ADAPT:RI for 30–60 min. (E) Fixed 1-mm brain slice from a LysMCre;tdTomatofl/fl mouse before ADAPT-3D processing. (F) The same section after 6 h of ADAPT: DC followed by 5 h of ADAPT:RI but without delipidation or (G) another section treated the same except for the addition of a 3-hour incubation in ADAPT:PDL before viewing immersed in ADAPT:RI. (H) Tiled confocal image of a single plane from a 1 mm section with a LysMCre;tdTomatofl/fl reporter (red) and nuclei stained with anti-histone antibody (cyan). (I) Confocal acquired fluorescent z-stacks of the brain stem and (J) cerebral cortex from the 1 mm section shown in F without ADAPT:PDL treatment (left) or from the 1 mm section shown in G with 3 h of ADAPT:PDL treatment (right). (K) Before and after top-down stereoscope image of fixed whole mouse brain and corresponding quantification of the brain area for duplicate samples following the iDISCO + method including 4 h of RIM in ethyl cinnamate or (L) following ADAPT-3D tissue processing consisting of 48 h ADAPT:DC, 36 h ADAPT:PDL, and 4 h ADAPT:RI. Dashed lines on graph in K and L link paired samples before and after. ** in J indicates the location of the corpus callosum.
