This study investigates the effects of autoimmune dry eye disease in a rat model using fluorescein dye to assess corneal damage. The methodology involves applying fluorescein to the cornea and analyzing the resulting fluorescence to evaluate the extent of damage to the corneal epithelium and Bowman's membrane.
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.
Use fluorescein scanning to measure corneal damage by adding 2 microliters of 0.2% fluorescein to the rat cornea. With a gloved finger, passively open and close the rat eyelids three times to spread the fluorescein dye on the surface of the eye.
After 1 minute, use a 3-milliliter syringe to draw 1 milliliter of saline. Then, position the syringe about 2 to 3 millimeters anterior to the cornea, and gently apply saline onto the eye. With the background lights switched off, acquire images under the cobalt blue filter of an ocular imaging microscope.
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