Corneoscleral Button Isolation: A Surgical Procedure to Obtain Corneoscleral Button from Porcine Eye

Use sterile forceps to transfer the eyeball to a Petri dish. Remove the conjunctiva and muscle tissue around the eyeball using a scalpel blade # 15 and forceps. Then, while holding the optic nerve with the forceps, gently lift the eyeball, and transfer it to a half-liter jar filled with sterile PBS.

Once all eyeballs are cleared of surrounding tissue, use sterile forceps to move them to another half-liter jar filled with 3% povidone-iodine in PBS. After the eyeballs have been in the jar for one minute, transfer them to a third jar containing sterile PBS.

Place an eyeball on a clean Petri dish and use the forceps to hold it steady. With a # 10A scalpel blade, make a cut near the cornea. Use scissors to excise the cornea, leaving about 3 millimeters of sclera surrounding it. Ensure the sharp end of the scissors is in the supra-choroidal space and does not pierce the iris.

Hold the corneoscleral button with forceps, and use another pair of pointed-end forceps to gently separate the uveal tissue. Then, lift the corneoscleral button to separate it from the remainder of the eyeball. Briefly rinse the button in a 1.5% solution of povidone-iodine in PBS. Then, place the button in sterile PBS in a 12-well plate.