Intravitreal Injection Protocol for Mouse

Intravitreal injections were performed according to a published protocol (Qian et al., 2008 (link)). All procedures were performed under dim-red light. Dark-adapted mice were anesthetized by intraperitoneal injection of ketamine (100 mg/kg) and xylazine (6 mg/kg), and one drop of 0.5% tetracaine topical anesthetic was applied to the cornea. One microliter of the drug (MFA, 200 μM) was injected into the mouse vitreous body. Injections were done through the sclera on the nasal side of the eye approximately 1 mm posterior to the limbus with a 10-μl Nanofil syringe with a removable 35-gauge needle (World Precision Instruments, Inc., Sarasota, FL, USA). Intravitreal concentrations of MFA were estimated to be 10 μM by assuming a complete mixture and the vitreal volume to be ~20 μl for a mouse eye (Wang et al., 2015 (link)). Phosphate buffer saline (PBS) was used as an injection control. ERG recordings were performed 2 h after injection with animals kept in dark.