Institutional review board approval and written informed consent were obtained. The purpose of this study was to prospectively validate usefulness of diffusion-tensor (DT) fiber tractography of the corticospinal tract at 3-T magnetic resonance imaging, in combination with the subcortical motor-evoked potential (MEP) technique, as a tool for tractography-guided neurosurgery. DT imaging and corticospinal tractography were performed at 3 T in eight patients (four men, four women; mean age, 41 years; age range, 23-58 years) with intracranial space-occupying lesions. Tractography data were transferred to a neuronavigation system, and tractography-guided neurosurgery was performed. During lesion resection, subcortical MEPs were recorded. Positive MEP response was observed in four patients. No patients developed new motor weakness postoperatively. Complementary use of tractography and MEP may be useful for intraoperative depiction of corticospinal tracts.