Figure 5.

3D volumetric analysis of wildtype and Ndp^-/Yretinal and choroidal vasculature in cleared eyes

(A) 3D reconstruction and 2D optical cross sections of dispase treated, depigmented, cleared eyes from C57BL/6J and Ndp^-/Y stained with anti-CD31 antibodies to detect endothelial cells. The choroidal (red) has been manually segmented from the inner retinal (green) vessels and pseudo-colored.

(B and C) (B) High magnification 3D reconstruction and 2D optical cross sections of CD31 stained retinal vessels in 1-month old C57BL/6J and Ndp^−/Y retinas. Images were taken as ~1mm area centered at the optic nerve. The choroidal (red) has been manually segmented from the inner retinal (green) vessels and pseudo-colored. Hypovascularization of the inner retina is evident at (A) low and (B) high magnifications. Semi-automatically derived 3D vessel reconstruction of the CD31 stain accurately models the fluorescent CD31 signal. The 3D vessel construction allows for the (C) volumetric comparison of choroidal and inner retinal vessels in both C57BL/6J and Ndp^−/Y retinas. Scale bars = 500 μm. Data are represented as means ± SEM, two-way ANOVA with Tukey's post-hoc multiple comparisons test, ∗∗ denotes p < 0.01, p = 0.0043; n= 4 eyes (C57BL/6J) and 3 eyes (Ndp^−/Y) with vessel (choroid and inner) and genotype (C57BL/6J and Ndp^−/Y) as factors.