Objective. To explore the effect of 3D Slicer preoperative planning and intraoperative guidance with mobile phone virtual reality (VR) technology on brain glioma surgery. Methods. By means of retrospective study, the data of 77 brain glioma patients treated in the neurosurgery departments at The Second Affiliated Hospital of Wannan Medical College and Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2015 to January 2022 were analyzed, and the patients were divided into the experimental group (EG, n = 38) and the control group (CG, n = 39) according to the surgical modalities. Before surgery, all patients received positron emission tomography-computed tomography (PET/CT) scanning and magnetic resonance imaging (MRI) examination. For patients in EG, the DICOM format images acquired from PET-CT and MRI examinations were imported with the 3D Slicer software for 3D visual fusion reconstruction, acquiring VR images, and developing detailed preoperative planning. Then, the reconstructed images were imported into the Sina software on a mobile phone, and the surgery was performed with the assistance of VR technology; for patients in CG, traditional 2D images were used for tumor contour drawing by the subjective visual method, and the craniotomy was performed under a traditional microscope. Patients’ surgery indicators and Karnofsky Performance Scale (KPS) scores were compared between the two groups. Results. The number of cases with total resection, rate of total resection, hospital stay after surgery, and surgery time were significantly better in EG than in CG ( P < 0.05 ); after treatment, the KPS score was significantly higher in EG than in CG (75.66 ± 4.01 vs 65.36 ± 5.23, P < 0.001 ). Conclusion. Combining 3D Slicer preoperative planning with intraoperative mobile phone VR technology can promote the accuracy of brain glioma surgery, which is conducive to effectively removing tumors while protecting patients’ neural function.