Immunohistochemical sections were imaged on a Zeiss Imager Z.2 fluorescence microscope, equipped with ApoTome 2, an Axiocam 506 mono camera and an HXP-120V fluorescent lamp (Carl Zeiss Microscopy, Oberkochen, Germany). The ZEN 3.3 (blue edition) software (Carl Zeiss Microscopy) captured z-stack images using 20x and 40x magnifications. The quantification of the rod/cone OS length, cone density and the number of outer nuclear layers (ONLs) was done by averaging measurements from at least four sections per animal. Per section, three distinct measurements were taken and averaged (Roche et al., 2016 (link)). Dorsal and ventral sections taken approx. 300 μm from the center of the visual streak were considered as peripheral retina. Figures were prepared using Photoshop CS5 (Adobe, San Jose, CA, USA). OCT reflectivity profiles between the upper ganglion cell layer and the bottom retinal pigment epithelium (RPE) layer were generated and analyzed using the ImageJ software package (NIH, Bethesda, MD, USA) as described previously (Garcia Garrido et al., 2014 (link)).
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