Phenylephrine hydrochloride eye drops

Animals were anaesthetized by intraperitoneal injection of xylazine (10 mg/kg body weight) and ketamine (80 mg/kg body weight). Pupils were dilated with 1% tropicamide and 2.5% phenylephrine hydrochloride eye drops (Bausch & Lomb, Rochester, NY, USA) and a 6 mm coverslip positioned on gel lubricant (Viscotears, Novartis, Basel, Switzerland) was applied to the cornea. 1.5 µl of virus at 2.2 × 10¹² virus particles/ml was injected into the vitreous using a Hamilton syringe with a 10 mm 34-gauge needle (65N, Hamilton AG, Bonaduz, Switzerland) using a surgical microscope (M620 F20, Leica, Wetzlar, Germany). All animals received bilateral intravitreal injections of the same virus. Anaesthesia was reversed by intraperitoneal injection of atipamezole (10 mg/kg body weight). During recovery, 0.5% proxymetacaine hydrochloride and 0.5% chloramphenicol (Bausch & Lomb) was applied to injected eyes. All animals began testing 1 month after virus injection.

All experiments were approved by the local Animal Welfare and Ethical Review Body (University of Southampton). All experiments were performed in accordance with relevant guidelines and regulations, including the ARVO statement for the use of Animals in Ophthalmic and Vision Research. Reporting of animal experiments follows recommendations in the ARRIVE guidelines. All experiments in this study were performed on three-month old C57BL/6 J female mice (n = 22, including control mice). The mice were maintained in a standard 12 h dark/12 h light cycle, with food and water available ad libitum. Mice were administered reversible anaesthesia with a combination of ketamine (80 mg/kg, Chanelle) and xylazine (8 mg/kg, Bayer) in 0.1 mL of saline. Mouse pupils were maximally dilated using 1% tropicamide and 2.5% phenylephrine hydrochloride eye drops (Bausch & Lomb). The corneas of mice were kept hydrated throughout laser treatment and imaging procedures carried out with repeated topical application of artificial tears (Viscotears, Alcone). Anaesthesia was reversed at the conclusion of the procedure with 10% antisedan in sterile saline at 8 μl/g of mouse weight. For terminal procedures, mice were anesthetised with 0.1 ml pentobarbitol and trans-cardially perfused with 0.9% NaCl (Sigma-Aldrich, UK) containing 5 units/ml heparin sulfate (CP Pharmaceuticals, UK).