We have visualised the release and approximate diffusion profile of nitric oxide (NO) from activated murine macrophages using a high transmission microscope coupled to a high sensitivity photon counting camera. The images generated by NO were cell-associated and spread over an area of approximately 175 mu m from the activated macrophage. The signals obtained were dependent on the presence of exogenous L-arginine in the medium and followed a time course similar to that previously described for the generation of NO by the inducible form of NO synthase. The light signal was attenuated by the inhibitor of NO synthase, N-omega-nitro-L-arginine methyl ester. Studies using superoxide-deficient macrophages further confirmed that the signals detected were generated by NO rather than reactive oxygen intermediates. (C) 1996 Academic Press, Inc.