In order to explore the microstructure of bilateral cingulum bundles associated with side and handedness, fractional anisotropy was extracted from diffusion tensor imaging. Distinguished from the conventional region of interest-based method, a fiber-based analysis method called scale-invariant parameterization method was employed to ascertain the anisotropy along the cingulum bundle in 31 normal right-handers and 14 normal left-handers. The statistical results showed a remarkable left-greater-than-right asymmetry pattern of anisotropy in most segments of cingulum bundles except the most posterior segment, for both right-handers and left-handers. Interestingly, higher anisotropy of the right-hander than the left-hander was found in the bilateral cingulum bundles. No significant handedness-by-side interaction was obtained in the present study, however.