Staining (both haematoxylin and eosin and period acid–Schiff [PAS]) was performed as previously described11. Fixed intact eyeballs were dehydrated in an ethanol gradient, cleared in xylene, embedded in paraffin, and sectioned (5 μm thickness) along the sagittal plane. Corneal tissue morphology was observed by haematoxylin and eosin staining and assessed using an optical microscope; conjunctival goblet cell morphology was observed by PAS staining (as described below) and assessed using an optical microscope (Olympus Optical Co. Ltd., Tokyo, Japan). Ten to 15 paraffin sections at matching positions of the anterior segment were stained with a PAS Kit (Sigma-Aldrich, St. Louis, MO, USA) and counterstained with haematoxylin.

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