Before white-nose syndrome arrives in Nebraska, it is important to document the preexposure distributions of cave bats in the state. We examined the distributions of Myotis lucifugus (little brown myotis), Myotis septentrionalis (northern long-eared myotis), and Perimyotis subflavus (tri-colored bat) in eastern Nebraska by setting acoustic detectors for a single night at 105 sites in wooded habitats during summers of 2012 and 2014. We compared 2 methods of determining presence at each site. Results of our analyses are fine-scale distributional maps for these bats and some range extensions from published records. Results for M. septentrionalis and P. subflavus are largely consistent with previous reports. Results for M. lucifugus vary depending on the method of determining presence; however, our preferred method creates a pattern consistent with the known vouchered distribution of this species. The differences between published distributions of these species and distributions based on acoustic detection from our study might result from a lack of extensive netting in many areas of eastern Nebraska, underrepresentation of P. subflavus from mist net surveys in Nebraska, and a recent westward range expansion of P. subflavus and M. septentrionalis in southern Nebraska.