To investigate the values of diffusion tensor imaging (DTI) and virtual reality (VR) techniques in design surgery program of gliomas near eloquent regions. In this study, 35 cases were retrospectively analyzed with gliomas involved language areas or rolandic regions operated in Department of Neurosurgery, Peking University Third Hospital from January 2015 to January 2019. Surgery programs were performed by Dextroscope virtual reality system. The pre-operative data, such as the magnetic resonance imaging (MRI), magnetic resonance arteriography (MRA) and DTI was transferred into the VR computer for restitution,Tumors, neural fiber tracts and blood vessels were reconstructed to simulate operation and design individual surgical plan. Neurological function was evaluated 1 week, 1 month and 3 months after operation. Virtual reality three-dimensional images of the 35 cases were successfully achieved, including neural fiber tracts,blood vessels and the lesions. The displacement and destruction of fiber tracts, the anatomic relationship between tumor and important fiber bundle, artery and vein could be shown clearly. Surgical simulation and surgery program of VR of the 35 patients were successfully performed. The 3D images obtained from virtual reality near to the real surgery. Ten of the 35 cases were defined as rolandic regions tumors, 14 of the 35 cases were defined as language areas tumors and 11 of the 35 cases involved both language areas and rolandic regions. Complete resection of enhancing tumor (CRET) was achieved in 30 cases (85.7%), subtotal resection in 5 cases (14.3%), neurological function improved in 34 cases (97.1%) after operation,and 1 case had no improvement compared with that before(2.9%). Thirteen cases without neurological deficit pre-operation, showed transient neurological deficit ,which were recovered about 10 days post-operation, 12 of 22 cases with pre-operative neurologic deficit, improved one week postoperation, 9 of 22 cases with pre-operative neurologic deficit improved one month after operation, the rest 1 case was recurrent with glioblastoma with aggravated hemiplegia symptom after operation, who died of cerebral hernia 2 months later. Dextroscope virtual reality system can clearly expose and quantify the 3D anatomic relationship of tumors, neural fiber tracts and blood vessels surrounding gliomas near eloquent regions, which is helpful to design the best individualized surgery program, to improve surgical effect.