Magnetic resonance imaging (MRI) is a key technology that has been seeing increasing use in studying the structural and functional innerworkings of the brain. Analyzing the variability of brain connectome through tractography analysis has been used to increase our understanding of disease pathology in humans. However, there lacks standardization of analysis methods for small animals such as mice, and lacks scientific consensus in regard to accurate preprocessing strategies and atlas-based neuroinformatics for images. In addition, it is difficult to acquire high resolution images for mice due to how significantly smaller a mouse brain is compared to that of humans. In this study, we present an Allen Mouse Brain Atlas-based image data analysis pipeline for structural connectivity analysis involving structural region segmentation using mouse brain structural images and diffusion tensor images. Each analysis method enabled the analysis of mouse brain image data using reliable software that has already been verified with human and mouse image data. In addition, the pipeline presented in this study is optimized for users to efficiently process data by organizing functions necessary for mouse tractography among complex analysis processes and various functions.