Developing an understanding of factors that influence the accumulation and magnification of heavy metals in fish of the Laurentian Great Lakes is central to managing ecosystem and human health. We measured muscle tissue concentrations of heavy metals in Lake Michigan prey fish that vary in habitat use, diet, and trophic position, including alewife, bloater, deepwater sculpin, round goby, rainbow smelt, and slimy sculpin. For each individual, we measured tissue concentrations of four metals (chromium [Cr], copper [Cu], manganese [Mn], and total mercury [THg]), stable isotope ratios for trophic position (δ15N and δ13C), and individual fish attributes (length, mass). Total mercury concentration was positively related to total length and δ15N. Of all species, round goby had among the greatest increases in mercury per unit growth and was most isotopically distinct from other species. Profundal species (bloater, deepwater sculpin, slimy sculpin) had similar high THg tissue concentrations, possibly due to slower growth due to cold temperatures, whereas other species (alewife, round goby, rainbow smelt) showed more variation in THg. In contrast, other metals (Cr, Cu, Mn) had either a negative or no relationship to total length and δ15N, suggesting no bioaccumulation or biomagnification. Potential incorporation of mercury by sportfish may thus be related to species, age, diet, trophic position, and habitat of prey fish. Our findings serve as a foundation for understanding how heavy metals accumulate in Lake Michigan food webs and highlight the continued need for management of metal input and cycling in Lake Michigan.