We quantified and compared movement and microhabitat use of Eastern Copperheads (Agkistrodon contortrix) in fragmented and unfragmented habitats to determine the effects of fragmentation on movement and habitat use. We used thread bobbins to track movement and calculate straight-line distance (SLD) moved, total distance (TDM) moved, and occupied area for individual snakes. Microhabitat use was characterized by quantifying number of trees, woody vegetation stems, herbaceous vegetation stems, percent grass coverage, and percent canopy coverage at each location a snake was observed, and at an equal number of randomly selected locations. Neither SLD nor TDM differed between fragmented and unfragmented habitats. Overall average SLD moved was 24.1 m and TDM was 39.6 m over 48 h. Although SLD and TDM did not differ between sites, mean occupied area ± standard error was significantly greater at the unfragmented (2,310.9 ± 272.7 m) compared with the fragmented site (1,025.9 ± 314.9 m). Microhabitat features were similar between the fragmented and unfragmented sites, and herbaceous vegetation and high canopy cover were associated with locations where snakes were observed at both sites. It is likely that Eastern Copperheads can persist in a variety of habitats in the southeastern United States because their preferred microhabitat features are widely distributed and common in both fragmented and unfragmented environments, demonstrating that they retain characteristics of a habitat specialist within heterogeneous environments suitable for generalists.