This study was designed to investigate the correlation between S-100 protein serum measurements and neuroradiological findings in patients with head injury. We studied 278 patients with minor, moderate, and severe head injuries and 110 controls with no history of neurological disease. The study recruited patients from three Scandinavian neurotrauma centers. Serum levels of S-100 protein were measured at admittance, and computed tomographic scans of the brain were obtained within 24 h postinjury in all patients. In a subgroup of 45 patients with minor head injuries, magnetic resonance imaging was also performed. Increased serum level of S-100 protein was detected in 108 (39%) patients, and CT scan demonstrated intracranial pathology in 25 (9%) (brain contusion n = 13, subdural hematoma n = 6, epidural hematoma n = 2, traumatic subarachnoid hemorrhage n = 2, and brain edema n = 2). The proportion of patients with detectable serum level was significantly (p < 0.01) higher among those with intracranial pathology (92%) compared to those without (34%). The negative predictive value of an undetectable S-100 serum level was 0.99. Undetectable serum level of S-100 protein predicts normal intracranial findings on CT scan. Determination of S-100 protein in serum may be used to select patients for CT scanning.