Laser-Induced Choroidal Neovascularization in Mice

The L-CNV mouse model was performed as previously described [52 (link)]. Briefly, both eyes of 6-8 week old C57BL/6J mice were dilated using tropicamide, and subjected to laser treatment using 50 µm spot size, 50 ms duration and 250 mV pulses of an ophthalmic argon green laser wavelength 532 nm, coupled to a slit lamp. Three laser burns per eye were created around the optic nerve at 12, 3 and 9 o’ clock positions. OCT was performed in L-CNV mice as described previously [52 (link)], on days 7 and 14 post laser, using a Micron III intraocular imaging system (Phoenix Research Labs, Pleasanton, CA, USA). Briefly, eyes of anesthetised mice were dilated with 1% tropicamide solution (Alcon, Fort Worth, TX, USA) and lubricated with Gonak hypromellose ophthalmic solution (Akorn, Lake Forest, IL, USA). Horizontal and vertical OCT images were taken per lesion and L-CNV lesion volumes were obtained using the quantification method previously established [52 (link), 53 (link)]. To assess vascular leakage, FA was performed on day 14 post L-CNV by i.p. injection of 50 μl of 25% fluorescein sodium (Fisher Scientific, Pittsburgh, PA, USA). Fundus images were taken using the Micron III system and Streampix software.