Multimodal Imaging of Retinal Structure and Function

FP, SD-OCT, and FFA images were acquired sequentially using the Phoenix Micron III system (Tempe, AZ, USA), according to the manufacturer’s instructions. The mice were anesthetized with a mixture of ketamine and xylazine, followed by a topical administration of 0.125% atropine for pupil dilation. For improved visualization of the fundus, a topical drop of 2% Methocel^® (OmniVision, Switzerland) was administered before ophthalmoscopic observation. After SD-OCT scanning, an intraperitoneal injection of sodium fluorescein (10 mg/kg) was administered prior to FFA observation. To quantify structural changes, retinal thickness (or retinal sublayers) was automatically computed using the Insight software, as described previously [22 (link)]. The FFA images for the quantification analysis were saved in the 16-pixel format. After the removal of the fluorescence signal in the retinal vessels, the remaining areas (in terms of the number of affected pixels) are considered the leakage regions. The total leakage area was calculated using ImageJ (U.S. National Institutes of Health, Bethesda, MD, USA) [23 (link)].