Objective：To investigate the application of multimodal neuronavigation combined with intraoperative fluorescence guided technique in the high grade glioma of functional areas. Methods：Sixty patients were divided into the control group and the observation group. The control group received conventional craniotomy operation and the observation group operated by multimodal neuronavigation combined with intraoperative fluorescence guided technique. We collected tumor specimen at different distances and different staining region，and analyzed pathology and immunohistochemistry of these specimens. The total resection rate of tumor and nerve function disorder after operation was compared between two groups. The influence on prognosis of the patients was analyzed. Results：The observation group was successfully achieved preoperative function fusion and intraoperative navigation. Intraoperative fluorescence showed different staining regions. The pathological type of the two groups had no significantly difference. There was significant difference of high grade glioma resection rate and postoperative neurological function disorder and tumor recurrence of postoperative 3 mon between the two groups. Each index of immunohistochemical expressed significant changes in the trend at different distance of tumor specimens. The fluorescence staining area was consistent with the postoperative pathology. Conclusion：Multimodal neuronavigation combined with intraoperative fluorescence guided technique can improve the total resection rate of high grade glioma in brain function areas，and improve postoperative nerve function.