We characterized Indiana bat (Myotis sodalis) roosting habitat at three maternity colony sites in western North Carolina and eastern Tennessee. Using radio telemetry, we tracked six bats a total of 40 bat days (range 4–9 days/bat). In 1999, we located a primary roost in an eastern hemlock (Tsuga canadensis) snag (109 cm DBH) in the Nantahala National Forest, NC. In 2000, we located a primary roost in a pine (Pinus sp.) snag (39 cm DBH) in Great Smoky Mountains National Park (GSMNP), TN. Another primary roost was found in a pitch pine snag (P. rigida; 55 cm DBH) in GSMNP in 2001. Largest exit counts for the three colonies were 28, 23, and 81 bats. Primary roost sites were exposed to direct sunlight during most of the day. We also located six alternate roost trees: three pine snags, two red oak (Quercus rubra) snags, and one live sweet birch (Betula lenta). All three primary roosts located in this study were not used during subsequent summers. The eastern hemlock used in 1999 was still standing as of June 2001, while the two primary roosts in GSMNP had fallen within a year of being located. These records represent one of the first descriptions of Indiana bat maternity habitat in the southern United States.