(A) Representative endogenous TPEF 3D image reconstructions from healthy, LSIL, and HSIL cervical tissues. Individual TPEF images are created by the overlay of NAD(P)H and FAD fluorescence, which are pseudo-colored green and red, respectively. 3D reconstruction volumes correspond to healthy: 238 × 238 × 160 μm3/SIL: 238 × 238 × 120 μm3/HSIL: 238 × 238 × 60 μm3.
(B) Quantification of the average epithelial thickness detected from the healthy (blue; N = 4 participants; 10 tissue stacks), LSIL (light gray; N = 16 participants; 38 tissue stacks), and HSIL (dark gray; N = 5 participants; 15 tissue stacks) cervical tissues examined. Healthy versus SIL comparisons are also presented.
(C) Automated depth-dependent quantification of the intraepithelial differentiation gradient. Healthy, LSIL, and HSIL and healthy versus SIL comparisons are presented. Exact p values shown in panels, *significance at a = 0.05.
In (B) and (C), data are presented as quantile boxplots with median (white line) and 95% confidence diamond around the mean (gray diamond). Each point represents 1 optical image stack.