Surgical excision of brain tumors provides a means of cytoreduction and diagnosis while minimizing neurologic deficit and improving overall survival. Despite advances in functional and three-dimensional stereotactic navigation and intraoperative MRI, delineating tissue in real time with physiologic confirmation is challenging. Raman spectroscopy has potential to be an important modality in the intraoperative evaluation of tissue during surgical resection. In vitro experimental studies have shown that this technique can be used to differentiate normal brain tissue from tissue with infiltrating cancer cells and dense cancerous masses with high specificity, indicating the feasibility of this method for in vivo application.