Objective · To explore the feasibility of applying customized guide, based on digital image processing and 3D printing technology, to bone biopsy. Methods · Eleven patients (6 males and 5 females; 7-69 years old, average age of 36 years) diagnosed with bone tumor July 2017 to March 2018 in Shanghai Ninth Peoples Hospital were enrolled in this study. Routine CT and/or MRI data were taken as reference in designing the customized biopsy guide for 3D printingMedraw Print software. Needle insertion points were set under the supervision of surgeons to avoid damaging important anatomical structures such as nerves and blood vessels. 3D printed customized guide was designed to match local surface. The success of the puncture was confirmedX-ray or CT, and the accuracy of the puncture results was evaluatedthe pathological results of resection operation as a reference standard. The complications were analyzed. Results · The 11 patients included 4 cases with neoplasms in femur, 3 cases in pelvis, 2 cases in tibia, 1 case in scapula and 1 case in metacarpal. The needling positions in operation were consistent with the preoperative virtual design, and the guide plates fitted perfectly to the body surface. The results of biopsy of 8 patients (88.9%) matched with the postoperative pathology, while another 2 patients did not receive bone tumor resection becaof the benign biopsy. No complications were observed during or after the procedure. Conclusion · Customized guidedigital image processing and 3D printing may have a great value in improving the accuracy of preoperative tumor diagnosis and the effectiveness of its treatment.