The authors first divided the midsagittal corpus callosum of 22 healthy volunteers using three different subdivision methods: Witelson's geometric subdivision, boundary model-based subdivision, and medial model-based subdivision. Next, subdivisions of the corpus callosum were clustered on the basis of homogeneity of the fractional anisotropy values as produced from diffusion tensor images. Nine subregions of corpus callosum, similar in location and size, were successfully clustered when medial model-based or boundary model-based subdivisions were applied. These nine subregions of the corpus callosum were different from the seven subregions defined by Witelson's geometric subdivision, especially at the anterior and posterior tips of the corpus callosum. Our method of corpus callosum division can be potentially utilized to assess structural changes in the midsagittal corpus callosum.