A Technique to Induce Chronic Experimental Autoimmune Dry Eye in Rats

On day zero, distribute 200 microliters of the emulsion which contains 1 milligram of lacrimal gland extract and 200 micrograms of ovalbumin, and complete Freund's adjuvant into 1-milliliter Luer-lock syringes, and equip the syringes with 27-gauge needles.

Without using anesthesia, subcutaneously inject the emulsion at the base of the rat tails. Then, on day 14 in a similar manner, inject 200 microliters of the same antigen and incomplete Freund's Adjuvant. On the same day, intraperitoneally inject 300 nanograms of pertussis toxin in 100 microliters of PBS.

On day 48, weigh the required amount of ovalbumin and dissolve it in PBS, then, combine ovalbumin with the defrosted lacrimal gland extract prepared earlier in this video, making antigen solution containing 1 milligram per milliliter of ovalbumin and 1 milligram per milliliter of lacrimal gland extract.

After anesthetizing the rats, inject 5 microliters of antigen into the fornicial subconjuctiva of the eye, then, inject 20 microliters of antigen into the lacrimal gland.