The present study demonstrates narrowband short-wavelengths radiation- (400, 420, and 435.8 nm) induced cellular damage of cultured human retinal pigment epithelial cells using in vitro biological assays to determine wavelengths that are responsible for photochemical lesions of the retina. This work involved the exposure of retinal pigment epithelial (RPE) cells (ARPE-19) to narrowband light of three different wavelengths (400, 420, and 435.8 nm) using a xenon arc lamp and interference filters. Cellular viability, mitochondrial distribution, and nucleic acid (both DNA and RNA) damage were quantified after various energy levels of exposure, using the Alamar blue assay, and confocal laser scanning microscopy with two fluorescent stains (Rhodamine 123 and Acridine Orange). The results clearly show that 400 nm light radiation can cause significant dose-dependent decreases in RPE cell viability as well as degradations of DNA/RNA and mitochondria in RPE cells, while 420 and 435.8 nm light radiation cause no cellular damage. While further evaluations may be needed to assess specificity and confounding factors of these assessment tools, the results may be a matter for consideration in future IOL design efforts.