OBJECTIVE To investigate the histological diagnostic efficacy and accuracy of stereotactic biopsy for intracranial deep lesions under computerized tomography (CT) and magnetic resonance imaging (MRI) guidance. METHODS CT or MRI-guided stereotactic biopsy was performed in 605 cases with intracranial deep lesions. All the procedures were done under local anesthesia with Leksell stereotactic system. CT or MRI film was used to determine the coordinate in 450 cases and the computer-assisted planning for neurosurgery (CAPN) in 155 cases. RESULTS Brain tumors, inflammatory lesions, and abnormal but nonspecific lesions were diagnosed pathologically in 537 cases (88.75%), 30 cases (4.96%), and 13 cases (2.15%) respectively. In 20 cases (3.31%) the biopsy procedure failed to provide useful pathological evidence. The total positive rate of biopsy was 96.69%. Complications associated with biopsy occurred in 18 patients (2.98%), and 2 patients (0.33%) died following the procedure. CONCLUSIONS Stereotactic biopsy using MRI-guided stereotactic approach or CT-guided technique is a safe and reliable method for histological diagnosis of intracranial deep lesions.