To specify the spectral sensitivity of the retinal pigment epithelium (RPE) for blue light damage, pigmented rabbits were exposed to light of 408, 418, 439, 455, 485, and 500 nm (half-peak bandwidth approximately 12 nm). The range of radiant exposure was 15–275 J cm−2 (1.7–19 mW cm−2 for 0.5–5 h). Vitreous fluorophotometry was used to functionally evaluate the blood-retinal barrier at the RPE in vivo, and electron microscopy to visualize RPE ultrastructure in vitro. A significant increase in permeability of the blood-retinal barrier was seen only after exposure to light of 418 nm. Radiant exposure at threshold for permeability increase was 18 J cm−2. Electron microscopy of the RPE demonstrated dispersion and clumping of melanin granules. The results suggest that the RPE is most sensitive to light in the range 412–425 nm, possibly due to damage-mediating chromophores such as cytochromec oxidase and lipofuscin.