The 3-D vascular network highlights the spatial variations in microvascular obliteration that occurred in the secondary plexus and vertical sprouts following hyperoxia-induced injury in P12 mice. 3-D rendering of the retinal vasculature was performed from the (A) normoxia and (B) OIR intact retinas, revealing statistically significant obliteration of microvasculature in the OIR retina in P12 mice. (C) 3-D filament tracing of the vasculature was performed in the OIR group. (D-E) The results of 3-D rendering were compared between the two volumes of interest (VOIs) in the normoxia (D) vs. OIR (E) groups, demonstrating the absence of secondary plexus and vertical sprouts in the OIR mice. (F) Immunofluorescence images of flat-mount retinas only captured the phenotype in the primary plexus. (G) Quantification of the morphological and topological parameters revealed the statistically significant reduction in total vascular lengths, total volumes, connectivities and branching points in the OIR P12 mice (*p < 0.05, ** p < 0.01 *** p < 0.001, by unpaired two-tailed Student's t-test, n = 5 per group). Scale bar: 500 µm for A-C; 100 µm for D-E, 500 µm for F.