Using confocal imaging and fluorescent calcium indicators, light-induced elevation of intracellular Ca2+ concentration ([Ca2+]i) in Limulus ventral photoreceptors was shown to be initiated within 4 microns of the light-sensitive plasma membrane. Within 500 ms, elevation of [Ca2+]i spread throughout the light-sensitive rhabdomeral lobe of the photoreceptor, but barely penetrated the arhabdomeral lobe. During saturating illumination of measurement spots near the plasma membrane, [Ca2+]i rose at rates of 1-2 mM/s after a latent period of 14-40 ms, reaching peak concentrations of approximately 150 microM. Rapid elevation of [Ca2+]i persisted in the absence of extracellular Ca2+ and was therefore ascribed to release from intracellular stores. The elevation of [Ca2+]i was always detectable within 5 ms of the electrical response of the photoreceptor to light. In 14 out of 54 measurements, detection of elevated calcium preceded the electrical response. Cyclopiazonic acid, an inhibitor of endoplasmic reticulum Ca-pumps, greatly reduced the elevation of [Ca2+]i during bright flashes and the sensitivity of the electrical response to dim flashes. However, the maximal response to bright flashes was not diminished. Therefore, although the calcium release that we detect may be fast enough to contribute to the electrical response we are unable to demonstrate that it is absolutely required.