This article details the dissection technique for obtaining an intact retinal cup from the murine eye. The process involves careful removal of the eye layers to preserve the retinal structure for further applications.
The murine eye is covered by three layers: the outermost fibrous sclera, the middle choroid coat, and the innermost retina. The cup-shaped cavity between the retina and eye lens contains vitreous humor — a gelatinous medium that helps transmit light to the retina. In turn, the retina passes the resulting visual information via the optic nerve to the brain.
To extract the retinal cup, take a euthanized mouse and enucleate its eye. Submerge the eye in a Petri dish containing an appropriate ice-cold buffer to prevent tissue degradation. Place the Petri dish under a dissection microscope.
Snip off the remnants of the optic nerve from the base of the eyeball. Make a small incision at the junction of the retina and the cornea — a transparent covering around the eye lens. Using this opening, make a complete circular cut along the retina-cornea junction to detach the lens and the vitreous humor gel from the eye.
Remove the lens and vitreous humor to obtain a semi-circular eye cup containing the retina lined by choroid and sclera. Create multiple superficial cuts throughout the scleral portion of the eye cup. Use these cuts to separate the outer eye layers for obtaining an intact retinal cup for downstream applications.
Enucleate the eyes from an adult euthanized mouse, and place them into ice-cold PBS in a Petri dish, then, place the Petri dish under a dissection microscope. Using micro scissors, cut off the optic nerve, then, carefully remove the extra rectus muscles attached outside the eyeball.
Using a 30-gauge needle, punch a hole at the edge of the cornea. Then, use fine dissection micro scissors to make a circular cut along the edge of the cornea. Using sharp dissection forceps, remove the cornea, lens, and the vitreous humor from the eye cup. Make several small cuts on the scleral layer at the rim of the eye cup using fine dissection micro scissors. Use two sharp dissection forceps to hold onto the scleral tissue at each side, and pull on the scleral layer very carefully to remove it from the neural retina. Repeat this around the eye cup until all sclera is removed, and an intact retinal cup is obtained.
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