Automated Assessment of Retinal Vascular Integrity Across Species.

Purpose: To develop an automated approach for the analysis of fundus fluorescein angiography (FFA) images and apply this methodology to examine inner blood-retina barrier (iBRB) circadian integrity across species and age. Methods: We acquired and quantified retinal images from mouse, non-human primate (NHP), and human eyes taken at the morning and in the evening. To facilitate the analysis, we developed and validated the Fluorescent Ocular Vasculature Analysis Suite (FOVAS), allowing for the quantification of large temporal retinal FFA datasets, with automated analysis, graphing, and reporting. We then compared fluorescein intensity at various microvasculature regions and ages and examined fluorescein signal at distinct phases along a 10-minute imaging session. Results: We detected a significant circadian-related difference in iBRB leakage between the morning and the evening in young adult mice, NHPs, and humans, using the validated FOVAS to quantify the FFA images. This difference was reduced with age across species. Conclusions: FOVAS successfully quantified FFA images demonstrating high sensitivity for the detection of changes in vascular integrity across species and time.