Background: Since cone-beam computed tomography (CBCT) technology has been widely adopted in orthodontics, multiple attempts have been made to devise techniques for mandibular segmentation and 3D superimposition. Unfortunately, as the software utilized in these methods are not specifically designed for orthodontics, complex procedures are often necessary to analyze each case. Thus, this study aimed to establish an orthodontist-friendly protocol for segmenting the mandible from CBCT images that maintains access to the internal anatomic structures. Methods: The “sculpting tool” in the Dolphin 3D Imaging software was used for segmentation. The segmented mandible images were saved as STL files for volume matching in the 3D Slicer to validate the repeatability of the current protocol and were exported as DICOM files for internal structure analysis and voxel-based superimposition. Results: The mandibles of all tested CBCT datasets were successfully segmented. The volume matching analysis showed high consistency between two independent segmentations for each mandible. The intraclass correlation coefficient (ICC) analysis on 20 additional CBCT mandibular segmentations further demonstrated the high consistency of the current protocol. Moreover, all of the anatomical structures for superimposition identified by the American Board of Orthodontics were found in the voxel-based superimposition, demonstrating the ability to conduct precise internal structure analyses with the segmented images. Conclusion: An efficient and precise protocol to segment the mandible while retaining access to the internal structures was developed on the basis of CBCT images. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.