Abstract—We used I‐m2throw traps to examine habitat use by smallsized fishes within a mosaic of wet prairies and sloughs in the headwaters of the St. Johns River, Florida between August 1992 and November 1995. Estimates of total fish density and biomass varied temporally, but did not differ significantly between habitats. Patterns of habitat use, however, differed among the five numerically dominant species. Bluefin killifish, mosquitofish, and golden topminnows were more abundant in sloughs than in wet prairies. In contrast, Everglades pygmy sunfish were more abundant in wet prairies than in sloughs. Finally, the abundance of least killifish did not differ between habitats. Fish densities were positively correlated with plant biomass (i. e., habitat complexity) and negatively correlated with water depth (i. e., hydrology). Species richness and composition were similar among habitats. However, consistent differences in the relative abundance of numerically dominant species between habitats indicated some degree of habitat‐specific assemblage structure. Most species were concentrated into deeper sloughs during drying events. This assemblage of small‐sized fishes appears to respond relatively rapidly to changes in habitat structure and hydrologic conditions. We therefore recommend that resource managers consider using fishes as indicator taxa to evaluate the efficacy of ongoing restoration and management efforts in wetland systems.