Delivery of a Fluorescent Dye into the Subretinal Space of a Mouse

Take an anesthetized mouse and trim the whiskers to facilitate eye visualization.

Place the mouse under a microscope on a warming pad to maintain body temperature.

Dilate the pupil to enable retina visualization, then lubricate the eye to prevent dryness.

Pinch the conjunctiva, a transparent membrane, and make an incision.

Incise the underlying Tenon’s capsule and remove the connective tissue layer while rotating the eye.

This exposes the sclera, the outer layer of the eye, and provides access to the injection site near the optic nerve.

Make a scleral incision and insert a syringe containing a fluorescent dye into the subretinal space between the retinal photoreceptor layer and retinal pigment epithelium.

Inject the dye to create a controlled retinal detachment, then withdraw the needle.

Rinse the eye and apply an antibiotic to prevent infection.

Allow the mouse to recover and administer pain relief. The mouse is ready for imaging to confirm successful subretinal injection.

To begin this procedure, trim the whiskers of an anesthetized mouse to facilitate visualization and maintain the animal's body temperature at 37 degrees Celsius with a circulating water pad. Next, dilate its pupils with