Information regarding habitat types selected by federally endangered Florida panthers (Puma concolor coryi) at natal den sites is needed because habitats used for parturition and kitten rearing potentially influence fitness of individuals and viability of the population. We located 51 natal den sites of 30 female panthers and performed a Euclidean distance analysis to determine which habitat types were selected and avoided at dens in south Florida, USA. Panther dens were closer to upland hardwoods, pinelands, and mixed wet forests (P ≤ 0.003) and farther from freshwater marsh–wet prairie (P = 0.009). We recommend that habitat protection efforts prioritize blocks of land that have abundant patches of upland hardwood, pinelands, and mixed wet forests to maintain preferred panther denning habitat.