The success of autologous tooth transplantation depends on the activity of the periodontal ligament of the donor tooth. Its activity decreases with a longer exposure time. In order to reduce the exposure time of the donor tooth and quickly prepare the alveolar fossa highly matching with the donor tooth root, the clinical data of cone-beam CT (CBCT) was imported into Mimics 21.0 software in this study to obtain three-dimensional (3D) images of the jaw tissue structure. The images were used to extract the target area and select the target tooth. By analysing the information of the recipient region and the donor region, 3-Matic 13.0 software was used to implant the donor virtual into the target region, and restrictive personalised guidance was designed. Subsequently, the surgical guide template was printed by 3D printing technology, and the alveolar fossa was rapidly prepared in vitro. After the operation, the donor tooth was matched with the complete alveolar cavity using 3-Matic13.0 software. The depth deviation of the prepared alveolar cavity was measured within 2.0 mm, and the width deviation was about 1.0 mm. The maximum width deviation is 2.88 mm due to the tilt of the roots. With a high matching degree to the donor tooth root, it needs further and larger clinical studies. Key Words: CBCT, Tooth autotransplantation, 3D printing technology, Computer-aided imaging, Periodontal ligament.