4.4. Fluorescein Angiography

Mice with laser induction of CNV were treated with 20 mg/kg AN7 or saline (n = 6 in each group), administered IP immediately following laser application for a total of three times a week (on days 0, 2, 4 post CNV induction). On days 2 to 7 post CNV induction, mice were anesthetized, their pupils dilated, and 0.1 ml 2.5% fluorescein sodium (Novartis, Basel, Switzerland) was injected IP. Vascular leakage was evaluated using the Optos California UWF imaging system (Optos Inc., Marlborough, MA, USA). Color fundus images were also taken.

Two masked retina specialists, not involved in laser photocoagulation or FA imaging, interpreted the fluorescein angiograms. Each laser spot was classified to “Leakage”: Hyperfluorescence gradually increases in intensity over time, with the borders of hyperfluorescence becoming increasingly blurred on the late phases of imaging; or “Staining”: Hyperfluorescence gradually increases in intensity over time, but the borders remain fixed throughout the angiograms.