(A) Representative DIC, native autofluorescence (AF), filipin-stained free cholesterol (FC), and AF/FC-merged confocal photomicrographs of RPE cells from human BVMD (hBest1-T6R) and canine model (cBest1-R25X). Lipofuscin granules (green) and free cholesterol (filipin-positive puncta, cobalt blue) were highly associated in BVMD and cBest RPE cells. (B) Both the analysis of AF and FC emission profiles as well as Pearson correlation coefficient ‘r’ values indicated a strong positive correlation between endogenous lipofuscin fluorophores (FITC) and filipin-stained-FC in BVMD-T6R and canine cBest1-R25X RPE (r =0.85±0.03 and r =0.77±0.02, respectively). Scale bars: 10µm; magnified images: 5µm.