ABSTRACT. Fixed honey-bees were conditioned to a scent in a one-trial learning paradigm. In contrast to free-flying colour-conditioned bees, fixed scent-conditioned bees do not show a biphasic time dependence of the conditioned response. Small metal probes were used to cool localized areas of the antennal lobes, alpha-lobes, and calyces of the mushroom bodies of the brain at various times after conditioning. Localized cooling impaired the formation of memory in all three structures. The susceptibility to impairment after conditioning lasted approximately 3 min in the antennal lobes, 7 min in the alpha-lobes, and 10 min in the calyx area. It was possible to determine the influence of the contralateral hemisphere (relative to the learning antenna) by conditioning bees with only one antenna. No contralateral impairment was found in the antennal lobes; there were minor effects in the alphalobes; contralateral cooling led to reductions of the conditioned response only in the calyx area. The temperature dependence of memory impairment was different for the antennal lobes and the mushroom bodies (alpha-lobes and calyces). The latter were most sensitive to cooling at 5°C. No correlation between cooling duration and impairment of memory was found in the antennal lobes, but there was a linear relation between impairment and cooling duration in the alpha-lobes. Brief cooling (5 or 10 s) resulted in significant impairment of memory formation only in the calyx area. A series of control experiments proved that the impairment of memory is due to a reversible block of neural activity. It was possible to show that the impairment is specific for the three neural structures analysed, by cooling the lobula of the optic system at various times after conditioning. Lesions of the brain or application of KCl also resulted in time-dependent reductions of the conditioned response. Cooling the entire animal at various times after conditioning led to similar memory impairment to that resulting from localized cooling of the alphalobes.