Rat model of laser-induced choroidal NV

Male Brown Norway rats at 7–8 weeks of age had laser-induced rupture of Bruch’s membrane in 5 locations four disc areas from the optic nerve in each eye using a 532-nm diode laser photocoagulation (100 μm spot size, 100 ms duration, 150 mW power). Immediately after the laser, suprachoroidal injections of 30 ng or 300 ng of acriflavine were done in one eye and PBS in the fellow eye. The sclera was partially penetrated 2mm from the limbus with a 27-gauge needle and then a 33-gauge Hamilton syringe with a blunt end was inserted and using blunt dissection, the tip was advanced into the suprachoroidal space and 3 μl of acriflavine was injected. Fourteen days after injection, rats were euthanized, eyes were removed, retinas were removed, and eyecups were stained with 1:500 Alexa-594 labeled GSA. The area of choroidal NV at Bruch’s membrane rupture sites were measured by image analysis as described above. The mean of the 5 values in each eye was used as a single experimental value.