The oxygen dependence of hematoporphyrin derivative (HPD)-induced photoinactivation of Chinese hamster V79 cells was examined. Cells were treated with HPD (25 micrograms/ml) for 2 h and subsequently exposed to red light (greater than 590 nm) under either aerated or hypoxic (less than 10 ppm O2) conditions. Hypoxic cells were found to be extremely resistant to the lethal effects of HPD and light. The electron-affinic X-ray hypoxic cell sensitizer, SR-2508, did not sensitize hypoxic HPD-treated cells to light. The clinical implications of these findings are discussed, with consideration of the possibility that hypoxic areas in tumors may limit HPD phototherapy.