Gray squirrels (Sciurus carolinensis) are an important component of forest ecosystems in the eastern United States, but their ecology in subtropical habitats remains unstudied. This study was conducted on a population of gray squirrels in a 10 ha forest composed of hammock and upland pine habitats. The goals were to determine microhabitat selection, diet and seasonal activity patterns of squirrels in these two distinct habitats. Weekly walking surveys of the study site were conducted for 1 y. Gray squirrels showed a significant peak in activity during the fall (P < 0.001). Acorns were the primary food item and pine seeds (Pinus palustris, P. taeda) were of secondary importance. Use-availability analysis revealed that squirrels were observed significantly less often than expected in some tree species and significantly more often than expected in laurel oak (Quercus hemisphaerica), loblolly pine (P. taeda) and longleaf pine (P. palustris).