We analyzed intraspecific and sexually dimorphic osteological variation within the Baldwin County, GA, population of Didelphis virginiana (Virginia opossum). Where possible, 20 measurements (11 cranial/mandibular and 9 post-cranial) were obtained from each of 59 road-killed adult opossums (47 males, 12 females). Although range overlaps exist between the sexes for all measurements, males are significantly larger (P ≤ 0.05) for the following characteristics: greatest length of skull, condylobasal length, basal length, postpalatal length, nasal length, bicanine width, zygomatic breadth, length of mandible, scapula length, humerus length, radius length, femur length, and fibula length. Males exhibit significantly less (P ≤ 0.05) variation in mandibular tooth row length and significantly greater (P ≤ 0.05) variation in scapula length and fibula length. Intraspecific variation ranges from 14–54% for cranial/mandibular and 29–73% for postcranial measurements. Sexual dimorphism is most pronounced in cranial/mandibular dimensions. Comparisons with multi-state samples indicate that the continent-wide population of the Virginia opossum is very similar in terms of osteological dimension. The combination of low variation index and nonsignificant difference between the sexes in mandibular tooth row length suggests that this measure may be useful for distinguishing fossil opossum species.