The spectral characteristics of visual pigments are a major determinant in eliciting a response to light. To study the absorption maximum of the photoreceptors and their sensitivity to light in fish, rod outer segments (ROS) and cone cells were purified from the rock bream Oplegnathus fasciatus adapted to the dark. Ultraviolet/visible spectroscopic analyses of the ROS in the dark and its difference spectra indicated an absorption maximum of the visual pigment at ~ 500nm, and each eye of 1-year-old rock bream contained at least 1.2nmol of rhodopsin-like visual pigments. Microspectrophotometric analysis of the cone cell outer segments led to identification of three visual pigments with individual absorption maxima at 425, 520, and 585nm. Monochromatic light-emitting diode (LED) modules with different wavelengths (violet 405nm, blue 465nm, cyan 505nm, green 530nm, amber 590nm, and red 655nm) were constructed to examine the spectral sensitivity and photoresponse in association with the absorption maximum of the photoreceptor. Analysis of chromophore decay upon illumination with each LED at low (27μmol/m2/s) and high (343μmol/m2/s) intensities showed the highest sensitivity of the photoreceptor upon illumination with the 505-nm cyan LED, followed by LEDs with wavelengths of 530nm > 465nm > 405nm > 590nm > 655nm. Photoresponse analysis of the fish using a video tracking system, in the dark and upon illumination, also showed faster movement of fish with illumination with the cyan LED followed by in the order of green ≈ blue > violet > amber > red. These results indicated that a light with a wavelength closer to the absorption maximum of rhodopsin was more effective in eliciting a response to the light.