Abstract: Objecitve To explore the role of DTI-Tractography and intraoperative nuclei mapping in microsurgery of adult brainstem gliomas. Methods Nineteen patients with adult brainstem gliomas were recruited for this study in our hospital from 2010 to 2012. The role of DTI-Tractography and intraoperative nuclei mapping in the microsurgery was retrospectively reviewed. Results Two patients underwent almost total resection, while 11 patents underwent subtotal resection and 6 patients underwent partial resection. Neurological functions improved or remained stable in 14 patients(73.7%)after surgery; whereas, 5 patients(26.3%) experienced deteriorated or transitory new symptoms. Follow-up visits found that 17 patients(89.5%)had improved or stable neurological functions. Conclusion DTI-Tractography and intraoperative nuclei mapping provides great support to the choice of surgical approach and the determination of degree of resection. It helps achieve maximal safe resection of adult brainstem gliomas.