Viscotears liquid gel

Adult (6-8 weeks old) C57BL/6J mice (Harlan, Indianapolis, IN) were housed and bred in a Intravitreal injections were performed under a surgical microscope using mice that were anaesthetized with isoflurane (Merial; Animal Health Ltd., Essex, UK). Pupils were dilated using 1% tropicamide and 2.5% phenylephrine (Chauvin, Essex, UK). Viscotears Liquid Gel (Novartis Pharmaceuticals UK Ltd., Surrey, UK) were used to improve the visibility of the fundus. A 33gauge needle (Hamilton Bonaduz AG, Bonaduz, Switzerland) was inserted from the limbus with a 45° injection angle into the vitreous. The direction and location of the needle was monitored through the microscope. One μL of Pull-Et-RhB, Pull-Hy-RhB and Pull-Es-RhB at 500 ng/mL, or 1 μL of PBS, pH 7.4, were injected intravitreally using a repeating dispenser (PB-600-1; Hamilton Bonaduz). Fundus images were taken with the Micron IV system (Phoenix Research Labs, Pleasanton, CA, USA) at 6 and 12 hours after intravitreal injection. The mice received 100 µL of fluorescein concanavalin A (Vector Laboratories, Burlingame, CA, USA) intravenously to label the posterior segment blood vessels and 15 min later, the eyes were collected, retinal flat mounts prepared, and examined by confocal microscopy (Leica SP5, Leica Microsystems, Wetzlar, Germany).