Brain glioma invariably relapses since the tumor is difficult to remove completely. Accurate identification of tumor subtypes and tracing of tumor boundaries intraoperatively are the key clinical issues. Raman spectroscopy has a good application prospect in the intraoperative diagnosis of brain glioma due to its advantages of nondestructive, label-free, histology imaging and high specificity. In this article, the research and application of Raman spectroscopy in the in vitro diagnosis and intraoperative navigation-related diagnosis and treatment of brain glioma are reviewed.