Photosensitive pigments are formed in vitro from rat opsin with the rods in suspension or as digitonin solutions. Rhodopsin, isorhodopsin I, and isorhodopsin II were generated with 11-cis, 9-cis and 9, 13-dicis retinal, respectively. An isorhodopsin I analogue was formed on combination with 9-cis 13-desmethyl-14-methylretinal. The absorption spectra and photosensitivities of these pigments were found to be similar to the corresponding bovine pigments although the rates of formation are several fold slower.