Crayfish are ecologically important in freshwater systems worldwide and are imperiled in North America and globally. We sought to examine landscape- to local-scale environmental variables related to occupancy and detection probability of a suite of stream-dwelling crayfish species. We used a quantitative kickseine method to sample crayfish presence at 102 perennial stream sites with eight surveys per site. We modeled occupancy (psi) and detection probability (P) and local- and landscape-scale environmental covariates. We developed a set of a priori candidate models for each species and ranked models using (Q)AICc. Detection probabilities and occupancy estimates differed among crayfish species with Orconectes eupunctus, O. marchandi, and Cambarus hubbsi being relatively rare (psi < 0.20) with moderate (0.46–0.60) to high (0.81) detection probability and O. punctimanus and O. ozarkae being relatively common (psi > 0.60) with high detection probability (0.81). Detection probability was often related to local habitat variables current velocity, depth, or substrate size. Important environmental variables for crayfish occupancy were species dependent but were mainly landscape variables such as stream order, geology, slope, topography, and land use. Landscape variables strongly influenced crayfish occupancy and should be considered in future studies and conservation plans.