A robust standardized method for segmentation, quantification, and normalization of pediatric hippocampal volumes using magnetic resonance imaging is presented. The method will find application in time course measurements of hippocampal volumes in pediatric patients who suffer from temporal lobe epilepsy and was tested prospectively on six control patients (13-60 mo of age). The un-normalized hippocampal volumes obtained using our segmentation method ranged from 3.85 to 6.38 mL, in agreement with previously published results. Inter-and intraobserver variability of the segmentation method was determined to be 13.3% and 2.8%, respectively. Four different methods of volume normalization were tested. Normalization is required to adjust for age-related increases in hippocampal volume. The normalization approach that seemed to compensate best for growth-related hippocampal volume changes was based on a simple estimation of intracranial volumes. This is the first report of a consistent and reliable method for segmentation and normalization of hippocampi from pediatric patients that can be used to study the progression of neurologic diseases in children. Noninvasive neuroimaging techniques can be used to examine the structural and pathophysiologic bases underlying numerous disease processes. MRI is the imaging technique of choice for evaluating patients with epilepsy inasmuch as TLE often leads to anatomic lesions (1).The primary features of the mesial temporal sclerosis associated with TLE can be detected with MRI, and these features correlate well with known histologic features (2, 3). Hippocampal atrophy and increased signal on T 2 -weighted magnetic resonance (MR) images can be determined by qualitative visual analysis of MR images. Although visual analysis of hippocampal sclerosis can achieve a sensitivity of 98% and a specificity of 93%, the additional advantages of quantitative volume analysis include reliable lateralization of atrophy and increased objectivity of analysis (3). Absolute hippocampal volumes measured from fast spin-echo MR images have been shown to correlate with the hippocampal neuronal density of CA1, CA2, and CA3; such a correlation was absent with visual analysis of increased T 2 signal (4). Van Paesschen et al. (5) demonstrated that increased T 2 signal has a neuropathological basis different from that of hippocampal volume loss. Thus, hippocampal volumetric analysis can provide a reliable, noninvasive method for identification of hippocampal sclerosis from MRI.Visual analysis of MR images may detect gross hippocampal atrophy, but smaller degrees of volume loss may be overlooked and small asymmetries of shape may be misinterpreted as volume loss. Both histologic and volumetric studies have shown that hippocampal sclerosis in TLE tends to occur ipsilateral to the epileptogenic temporal lobe, but bilateral volume loss can also be present (4, 6, 7). Side-to-side comparison of hippocampal volumes can identify hippocampal sclerosis on the atrophic side, but absolute hippocampal volumes must be us...