Diffusion measurements were performed on the N-acetyl aspartate (NAA) signal in in situ brains (in vivo and post-mortem) and on in vitro brain tissue at 37 degrees C using wide ranges of b-values (from 0 up to 4.5 x 10(6) s/cm2 and 35.8 x 10(6) s/cm2 for the in vivo and the in vitro cases, respectively). In vivo and in vitro NAA signals attenuation due to diffusion was measured at fixed diffusion times (tD). In the in vitro cases the effect of tD on the apparent diffusion coefficients (ADCs) of NAA was evaluated. From these experiments the following observations and conclusions were made: (1) NAA signal attenuation both in vivo and in vitro is not mono-exponential and could be fitted by bi-exponential fitting function; (2) analysis of the low b-value range only (up to 0.5 x 10(6) s/cm2) gives a mono-exponential decay (r = 0.999); (3) in both cases the obtained ADCs are sensitive to the diffusion time; (4) the ADCs of the pre- and post-mortem cases are nearly similar; (5) the ADCs obtained from the bi-exponential fitting function decrease when the diffusion time increases; and (6) both the fast and the slow diffusing components of NAA show a considerable restriction by what seems to be a non-permeable barrier from which two compartments were identified, one having a size of 6-8 microns and the other of approximately 1-2 microns in size. It seems conceivable that the two populations identified in the diffusion experiments represent primarily the NAA in the cell body (soma) and in the neurital space (axons and proximal dendrites).