DTI has been used to noninvasively resolve major thalamic nuclei using an unsupervised clustering algorithm. However, rigorous validation of the method has not been studied. Here, we evaluated the method by comparing the parcellation results with histology in the same non-human primate. We found that some nuclei were clustered with larger differences from histology. That probably was because the diffusion properties of these nuclei were not coherent. In addition, the pipeline constructed in this study is also a framework to validate other approaches for thalamic nuclei parcellation.