The cold stress is considered the bottleneck problem limiting the development of red tilapia industry. Fish (initial weight: 72.71±1.32 g) was divided into the cold stressed group (C) and the control normal water temperature group (N). The water temperature was reduced by 2℃ per day from 20℃ to 8℃ in cold group, and the control group kept the water temperature at 20℃ during the experiment. At the end of the experiment, the gill(G)、liver(L) and skin(S) tissues from the fish of cold(C) group and control normal water temperature (N) group were taken. The DIA proteome analysis of 18 samples was completed, and 6341 trusted proteins containing quantitative information were obtained. Our aim is to understand the protein expression profiles of cold stress in red tilapia.