Eclipse e400 with a ds fi1

Eyes with ocular adnexa and extraorbital lacrimal glands were surgically excised, fixed in 10% formalin, and embedded in paraffin, and 8-µm sections were cut using a microtome (Microm HM 340E, Thermoscientific Wilmington, DE, USA). Sections were stained with haematoxylin and eosin for evaluation of morphology. Goblet cells in sections were stained with periodic acid-Schiff (PAS) reagent and were examined, photographed and counted with a microscope equipped with a digital camera (Eclipse E400 with a DS-Fi1; Nikon, Inc., Melville, NY, USA) as previously described [75 (link)]. The number of positively stained goblet cells in the superior and inferior conjunctiva was counted, and the length of the basement membrane between the first and last goblet cell was measured. The data are presented as the average number of goblet cells per millimeter per mouse.

Five right eyes from each sex, at 8W and 15M, were surgically excised, fixed in 10% formalin, paraffin embedded, and 8-μm sections were cut. Ocular sections were cut at the center of the eye, where the lens has its maximum diameter. Sections were stained with H&E for evaluating morphology and with periodic-Schiff (PAS) reagent for measuring goblet cell density. The superior and inferior conjunctiva were examined and photographed with a microscope equipped with a digital camera (Eclipse E400 with a DS-Fi1; Nikon, Melville, NY, USA). The number of goblet cells in the superior and inferior conjunctiva was measured in 3 sections from each eye that were at least 300 μm apart, using image-analysis software (NIS Elements Software, version 3.0, BR, Nikon) and expressed as number of goblet cells per mm as previously reported [21 (link)].