U cmad3 adapter

An Olympus BX50 Microscope (Model U-SD0, Olympus Optical Co., Ltd.) with an Olympus DP70 camera and Olympus U-CMAD3 adapter was used with the AnalySIS program for imaging. For enumeration of POU2F3⁺ cells, five images at x40 magnification were acquired for each tissue section, ensuring no overlap and samples scored blind. All epithelial cells were counted using Image J (22 (link)). Following counting of all POU2F3⁺ cells within the epithelial cells, the percentage of POU2F3⁺ cells/all epithelial cells was calculated. The mean percentage of POU2F3⁺ cells was calculated across the five images for each sheep.

Extraocular tissues were excised immediately after enucleation, to ensure proper penetration of the fixative. Then, the eyes underwent a two-step fixation process. For this purpose, a 10% formaldehyde solution was utilized, at ten times the volume of the tissue being studied. For the first 24 h, the ocular globes were submerged in the fixation solution, as a whole. Subsequently, they were briefly removed from the fixative, and sectioned in half, along the anatomical sagittal plane (with the blade placed perpendicular to the superior and inferior rectus muscles). Then, the obtained halves were fixed for a second 24 h period, in the same solution. Through employing a preliminary fixation of the entire globe, collapse upon halving was avoided. Ultimately, samples were embedded in paraffin and sectioned at 5 μ, then stained with haematoxylin-eosin and examined using an Olympus BX43F light microscope (Olympus, Tokyo, Japan). Pictures of representative areas were captured using the microscope mounted camera (Olympus UC30 camera with the Olympus U-CMAD3 adapter).