Traditionally studies of cold tolerance in insects have focused on seasonal adaptations related to overwintering that are observed after weeks or months of exposure to low temperature. In contrast, an extremely rapid cold-hardening response was observed in nonoverwintering stages that confers protection against injury due to cold shock at temperatures above the supercooling point. This response was observed in nondiapausing larvae and pharate adults of the flesh fly, Sarcophaga crassipalpis, nondiapausing adults of the elm leaf beetle, Xanthogalwruca luteola, and the milkweed bug, Oncopeltus fasciatus. The rapid hardening response is correlated with the accumulation of glycerol.