The tricolored bat (Perimyotis subflavus) once commonly occurred throughout the eastern and central United States, but is now experiencing range-wide population declines primarily due to white-nose syndrome and forest fragmentation. Conservation efforts for this species require more information regarding summer maternity roost characteristics, especially in the Ozark Highlands of Missouri, where the species has declined precipitously. Herein we report the capture and tracking of one female tricolored bat to two summer roosts in southeastern Missouri. Tree roosts differed by their substrates, as one was within a cluster of dead leaves on a snag and the other was located under exfoliating bark; otherwise, the roosts and surrounding vegetation were similar. Both roosts were located in large (>30 cm diameter breast height) oak (Quercus spp.) trees under a dense canopy of trees with similar basal areas. While our capture success was limited over two years, details regarding tricolored bat roost ecology are imperative for the species' conservation and management.