Significant attention has been paid to the adverse effects of dioxins, dioxin-like polychlorobiphenyls (DL-PCBs) and per-and polyfluoroalkyl substances (PFAS) in environment and food on ecosystem and human health. However, limited information exists regarding the occurrence, tissue distribution and exposure risk of these pollutants in crayfish. This study investigated tissue-specific distribution characteristics, influence factors and exposure risk of dioxins, DL-PCBs and PFAS in crayfish. Dioxins, DL-PCBs, and PFAS were detected in crayfish samples, with concentrations ranging from 0.361 to 1.91 pg TEQ g−1 for sum dioxins and DL-PCBs, and 1.54–21.2 ng g−1 for sum PFAS. The highest concentrations of dioxins, DL-PCBs TEQ and total PFAS were found in crayfish viscera and accounting for over 69% of the relative body burden (RBBs) of these pollutants. The specific-tissue distribution of these pollutants linked to tissue fat and protein content and log Kow and molecular weights of homologues. The consumption of crayfish edible tissue posed no health risks from exposure to dioxins and DL-PCBs as well as sum 4 PFAS in adults. However, consumption of crayfish viscera significantly increased exposure risk. This study can provide valuable insights into the tissue-specific distribution and human exposure risks associated with dioxins, DL-PCBs and PFAS in crayfish.