1. Confocal Ca2+ imaging was used to measure spontaneous release events (Ca2+ sparks) in fluo-3-loaded isolated rat ventricular myocytes. 2. The microscopic Ca2+ release flux underlying Ca2+ sparks was derived by adapting the methods used previously to describe macroscopic Ca2+ release from cell-averaged Ca2+ transients. 3. The magnitude of the local release fluxes varied from 2 to 5 μM ms-1, depending on SR Ca2+ loading conditions. Following spontaneous activation, the release flux rapidly decayed (τ = 6-12 ms). The rate of termination of release flux was found to be directly related to the magnitude of the flux (r2 = 0.88). 4. The rate of termination of local release flux was slowed in the presence of FK506, a compound that is known to reduce inactivation of SR Ca2+ channels in vitro. 5. These results suggest that termination of release flux during sparks is not due to a spontaneous stochastic decay process or local depletion of Ca2+ from the SR, but rather involves an active extinguishing mechanism such as Ca2+-dependent inactivation or adaptation.