Prepare either virus dilution with PBS prepare. Injection is up to 100 μL per mouse.

Take one mouse from the cage into an induction chamber and start isoflurane at 3% with flow rate of ~1L/min

After the mouse has slow breathing (~1/s) move the mouse to a nose cone delivery of isoflurane at 1.5% with around the same flow rate.

Expose the eye ball with two fingers (above and below) and make sure the you are not pressing on the trachea and the mouse's breathing is regular.

Use a 31 G insulin syringe at a 30-degree angle beveled downwards until you hit the bone, retract a bit and slowly inject. See cited paper for more details.

Equipment

Value	Label
U-100 BD Ultra-Fine™ Short Insulin Syringes	NAME
Syringe and neecdle	TYPE
BD Medical	BRAND
BD328438	SKU

Remove the needle slowly and press the eyelid shut for a few seconds.

Prior to recovery, apply 1-2 drops 0.5% ophthalmic Proparacaine to the injected eye