Detailed measurements of water diffusion within the prostate over an extended b-factor range were performed to assess whether the standard assumption of monoexponential signal decay is appropriate in this organ. From nine men undergoing prostate MR staging exams at 1.5 T, a single 10 mm thick axial slice was scanned with a line scan diffusion imaging (LSDI) sequence in which 14 equally spaced b-factors from 5 to 3500 s/mm 2 were sampled along three orthogonal diffusion sensitization directions in 6 minutes. Due to the combination of long scan time and limited volume coverage associated with the multi-b-factor, multi-directional sampling, the slice was chosen online from the available T2-weighted axial images with the specific goal of enabling the sampling of presumed noncancerous regions of interest (ROI's) within the central gland (CG) and peripheral zone (PZ). Histology from pre-scan biopsy (N = 9) and post-surgical resection (N = 4) was subsequently employed to help confirm that the ROIs sampled were non-cancerous. The CG ROIs were characterized from the T2-weighted images as primarily mixtures of glandular and stromal benign prostatic hyperplasia (BPH) which is prevalent in this population. The water signal decays with bfactor from all ROI's were clearly non-monoexponential and better served with bi-vs monoexponential fits, as tested using λ 2 based F-test analyses. Fits to biexponential decay functions yielded inter-subject fast diffusion component fractions on the order of 0.73 ± 0.08 for both CG and PZ ROIs, fast diffusion coefficients of 2.68 ± 0.39 and 2.52 ± 0.38 μm 2 /ms and slow diffusion coefficients of 0.44 ± 0.16 and 0.23 ± 0.16 um 2 /ms for CG and PZ ROI's, respectively. The difference between the slow diffusion coefficients within CG and PZ was statistically significant as assessed with a Mann-Whitney non-parametric test (P < 0.05). We conclude that a monoexponential model for water diffusion decay in prostate tissue is inadequate when a large range of b-factors is sampled and that biexponential analyses are better suited for characterizing prostate diffusion decay curves.