Diffuse axonal injury (DAI) is a common mechanism of traumatic brain injury (TBI) for which there is no well-accepted anatomic measures of injury severity. The present study aims to quantitatively assess DAI by measuring white matter lesion volume visible in fluid-attenuated inversion recovery (FLAIR) weighted images and to determine whether higher lesion volumes are associated with unfavorable functional outcome 6 months after injury. Twenty-four patients who experienced moderate to severe TBI without extra-axial or major cortical contusions were included in this study. Lesion volume was assessed by quantifying areas of hyperintensities in the white matter utilizing digitized FLAIR images. Two independent raters processed the magnetic resonance (MR) images and determined the total DAI volume. Functional outcome was assessed at 6 months after injury using the Glasgow Outcome Scale-Extended (GOSE). Interclass correlation analyses showed very high interrater reliability for each measure between the two raters (Interclass Correlation Coefficient = 0.95, p <or= 0.001). Total DAI volume was significantly, although modestly, correlated to GOSE (r = -0.453, p = 0.034). White matter lesion volume resulting from DAI can be quantitatively and reliably assessed from standard FLAIR-weighted MRIs. Patients with greater DAI volume have poorer functional outcomes. These methods may be useful in stratifying injury severity and for the assessment of DAI-directed therapies.