Abstract: The aim of this study was to determine the differences in the anatomical site of a gunshot entrance wound and the direction of the bullet path between right- and left-handed subjects who committed a suicide by a single gunshot injury to the head. The retrospective autopsy study was performed for a 10-year period, and it included selected cases of single suicidal gunshot head injury, committed by handguns. We considered only contact or near-contact wounds. The sample included 479 deceased, with average age 47.1 ± 19.1 years (range, 12–89 years): 432 males and 47 females, with 317 right-handed, 25 left-handed, and 137 subjects with unknown dominant hand. In our observed sample, most cases involved the right temple as the site of entrance gunshot wound (about 67%), followed by the mouth (16%), forehead (7%), left temple (6%), submental (2%), and parietal region (1%). The left temple, right temple, and forehead were the sites of the gunshot entrance wounds, which were the best predictors of the handedness of the deceased (Spearman ρ = 0.149, P = 0.006). Our study showed that the direction of the bullet intracranial path in cases of suicide was even a more potent predictor of the handedness of the deceased (Spearman ρ = 0.263, P = 0.000; Wald = 149.503, P = 0.000).