Diffusional anisotropy of human brain was investigated clinically in six adult volunteers, eight premature neonates, and three infants aged 5-10 months. Diffusion-weighted magnetic resonance imaging was performed with gradient b of 450 sec/mm2. The direction of diffusion-sensitive gradients was changed among x, y, and z axes according to the orientation of neurofibers in white matter. T1- and T2-weighted images also were obtained for evaluation of myelination. Diffusional anisotropy was demonstrated in white matter in the adults. Extensive signal attenuation was observed when gradients were parallel to white matter fibers. Conversely, in neonates, diffusional anisotropy of white matter, in which no myelination was shown on T1- and T2-weighted images, was weak. Diffusional anisotropy was more distinct after brain maturation, as observed in adult white matter. Detection of diffusional anisotropy is useful in evaluating neonatal brain development and various white matter disorders.