Using our custom-built confocal scanning laser retinal angiography system (50), in vivo fundus FA and ICGA of the CNV lesion were performed. Continuous-wave laser modules at 488 and 785 nm were used as excitation sources for fluorescein and ICG, respectively. A raster-scanning pattern of excitation lasers was achieved by a scanner system consisting of a rotating polygonal mirror (MC-5; Lincoln Laser) and a galvanometer-based scanning mirror (6230H; Cambridge Technology) and delivered to the back aperture of an imaging lens. A high–numerical aperture (NA) objective lens (PlanApo λ, NA 0.75; Nikon) was used as the imaging lens to provide wide-field fundus fluorescence images. Fluorescence signals detected by photomultiplier tubes (R9110; Hamamatsu Photonics) were digitized by a frame grabber and reconstructed to images with a size of 512 × 512 pixels per frame in real time. To visualize late-phase (6 min) FA and ICGA images using the angiography system, 10 mg of fluorescein sodium (Alcon) and 0.15 mg of ICG (Daiichi Pharmaceutical) were administered intraperitoneally and intravenously, respectively. The imaging procedure was performed under systemic anesthesia and pupil dilation to improve the quality of images. Leaky areas from CNV were calculated as the total measured hyperfluorescent areas in FA images divided by the total measured CNV areas in ICGA images using a Java-based imaging software (ImajeJ, NIH; available at http://rsb.info.nih.gov/ij).

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