Retinal Histology Analysis of Mutant Mice

Eyes were enucleated, fixed in glutaraldehyde (2.5% in cacodylate buffer) for 12-24 h at 4 ${}^{\circ }$ C, trimmed, post-fixed in 1% osmium tetroxide, and embedded in Epon 812 as described [77 (link)]. Retinal cross-sections of 0.5 $\mathrm{\mu }$ m were cut through the optic nerve head, stained with toluidine blue, and analyzed by light microscopy (Zeiss, Axioplan). OS and IS length, and the thickness of the ONL were measured at indicated distances from the optic nerve head using the Adobe Photoshop CS6 ruler tool (Adobe). Mann-Whitney nonparametric test was used to compare the overall OS and IS length, as well as ONL thickness of $ro{d}^{\Delta Vhl}$ and $ro{d}^{\Delta Cox10}$ mice to their respective controls. P-values < 0.05 were considered to show significant differences.