To begin, place an anesthetized mouse on a table. Apply a drop of ocular gel on both eyes. Place the animal in a head holder to secure its position.
Rotate the animal by 90 degrees so that the eye being operated on faces the ceiling. Secure the animal's vibrancy to clear the area around the eye. Secure the animal's body to minimize head movements caused by breathing.
Place a circular cover slip horizontally over the eye on top of the ocular gel. Adjust the animal's head as necessary to correctly position the cover slip. Add a drop of PBS on top of the cover slip, then carefully place the animal onto a microscope stage.
Adjust the objective turret of the microscope to the appropriate distance for observation. Activate the lasers to induce corneal nerve ablation. Once nerve ablation is complete, remove the animal from the head holder and place it in a cage on a heated plate.
The vortex was visibly absent one week after laser ablation. A group of immune cells were observed to have migrated to the illumination site. The stromal fiber bundles appeared to have thinned out.
