Fluorescein Staining in an Autoimmune Dry Eye Rat Model to Examine Corneal Damage

Start with an anesthetized rat presenting symptoms of autoimmune dry eye disease.

Apply an appropriate amount of fluorescein dye over the rat's cornea.

Close and open the rat's eyelid to ensure the fluorescein spreads evenly over the cornea.

Allow the dye to settle and adhere to the cornea.

In autoimmune dry eye disease, the immune cells mistakenly target and attack the lacrimal glands, decreasing tear production.

This leads to the desiccation of the corneal epithelium and damage to the underlying Bowman's membrane — an acellular zone anterior to the stroma.

The fluorescein molecules permeate through the damaged corneal barriers to reach the underlying collagen-rich stromal layer.

Here, the fluorescein molecules bind to collagen, resulting in the staining of the collagen fibers.

Using an appropriate imaging technique, acquire images of the stained cornea.

Analyze the number, area, and intensity of the green fluorescent spots to measure the extent of corneal damage.