The application of 3D printed individualized positioning guides in orthopedic clinical practice is a research hotspot and frontier in the international field of biomedical engineering. The current research difficulty lies in whether the nail tracks of the printed positioning guide model are consistent with those of the 3D simulation model. This project, based on the forefront of international research, uses CT scan images of sheep tibiae to establish three-dimensional models of tibial plateau fractures, plates, and guides, as well as 3D printed solid guide and plate models. These 3D printed guides are then applied in preoperative design and simulated surgical procedures in clinical orthopedic departments to achieve the treatment of posterior lateral tibial plateau fractures assisted by 3D printed individualized positioning guides. First, DICOM images of sheep tibiae are obtained through CT scanning to establish three-dimensional models of posterior lateral tibial plateau fractures and plate models, simulating different fracture conditions of the tibia and designing guides with nail tracks; next, 3D guide models and physical plate models are printed. The guide plate was fixed on different tibia specimens to simulate minimally invasive surgery, and screws were implanted according to the nail track. Then, CT scans and experimental verification were conducted to ensure that the position and length of the nail track were consistent with the parameters of the initial 3D simulation. Finally, this 3D printed guide plate technology was promoted to clinical orthopedic departments in hospitals. This project can be pilot applied at Gongli Hospital in Pudong New District, Shanghai, to achieve technological transformation. The transformed results can be widely used for preoperative design, simulated surgical procedures, medical teaching, and scientific research in clinical orthopedic departments of various hospitals. It serves as a bridge between virtual and real surgeries, improves the cure rate of orthopedic surgeries, and has significant economic and social benefits. It also holds important theoretical research significance, clinical value, application value, and broad market prospects.