The aim of this study was to examine the diffusive properties of adjacent muscles at rest, and to determine the relationship between diffusive and architectural properties, which are task-specific to muscles. The principle, second, and third eigenvalues, trace of the diffusion tensor, and two anisotropic parameters, ellipsoid eccentricity (e) and fractional anisotropy (FA), of various muscles in the human calf were calculated by diffusion tensor imaging (DTI). Linear correlations of the calculated parameters to the muscle physiological cross-sectional area (PCSA), which is proportional to maximum muscle force, were performed to ascertain any linear relation between muscle architecture and diffusivity. Images of the left calf were acquired from six healthy male volunteers. Seven muscles were investigated in this study. These comprised the soleus, lateral gastrocnemius, medial gastrocnemius, posterior tibialis, anterior tibialis, extensor digitorum longus, and peroneus longus. All data were presented as the mean and standard error of the mean (SEM). In general, differences in diffusive parameter values occurred primarily between functionally different muscles. A strong correlation was also found between PCSA and the third eigenvalue, e, and FA. A mathematical derivation revealed a linear relationship between PCSA and the third eigenvalue as a result of their dependence on the average radius of all fibers within a single muscle. These findings demonstrated the ability of DTI to differentiate between functionally different muscles in the same region of the body on the basis of their diffusive properties.