This article details the dissection and culturing of the anterior segment of the vertebrate eye, specifically using a porcine model. The procedure aims to facilitate the study of ocular disorders by isolating key structures within the eye.
The vertebrate eye is divided into the anterior segment - situated ahead of the lens - and the posterior segment - situated behind the lens. The anterior segment contains the cornea, aqueous humor, iris, ciliary body, and lens; while the posterior segment contains the vitreous humor, retina, choroid, and the optic nerve. The anterior segment can be cultured to study pathophysiology of ocular disorders.
To dissect the anterior segment from an animal model, begin with a freshly isolated porcine eye. Remove the extra-orbital tissue from the eye and wash with antibiotic-supplemented buffer. Place the eye over a sterile, antibiotic-soaked gauze to prevent contamination of the eye.
Using a scalpel, incise the eye near the equator region to open the corneoscleral shell - fibrous and vascular layer. Use curved scissors to isolate the anterior segment from the posterior eye portion. Carefully flip the anterior eye segment. Then, scoop out the vitreous humor and remove the lens.
Excise the iris radially towards the iris roots to reveal the trabecular meshwork - a pigmented tissue through which aqueous humor drains out of the eye. Remove the traces of the ciliary body, leaving a thin tissue band. Wipe the cornea with a wet cotton swab to remove the residual pigment. The anterior eye segment can now be cultured for further experiments.
After removing the extra orbital tissues, and washing the eyes with Antibiotic-Antimycotic-PBS, to hemisect the eye, place the eye on the Antibiotic-Antimycotic-PBS-soaked gauze. Use a sterile razor blade or scalpel to create an incision near the equator of the eye. Then, hemisect the eye using curved surgical scissors to isolate the anterior eye.
Use microscissors as a shovel to scoop the vitreous humor from the anterior segment, and remove the lens from the anterior segment with microscissors. Gradually, cut the iris to the iris root radially, until the trabecular meshwork is visible. Extend the cut 360 degrees around the iris, to expose the entire trabecular meshwork region, and clean up any remaining residual iris covering the trabecular meshwork, if necessary.
Trim the ciliary body remnants posterior to the trabecular meshwork region, leaving only a 1-millimeter thin band of the tissue. After placing the anterior segment in the petri dish, wet a cotton swab in the media, and gently dab in the center of the cornea to remove any pigment. Hold the eyes with forceps, and remove extra pigment around the sclera by wiping.
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