Appropriate therapy of brain stem lesions should be guided by an accurate diagnosis. Clinical evaluation combined with modern neuroimaging techniques may nowadays approach the diagnosis but not always with accuracy, thus leading to erroneous treatment. We report a series of 11 patients who underwent stereotactic biopsy for brain stem lesions. In 8 patients, the lesion was approached transfrontally. In the remaining 3 patients, the lesion was approached via the suboccipital transcerebellar route. There was no surgical mortality. Precise histological diagnosis was achieved in all patients. Three patients (27.2%) suffered complications attributed to the procedure and were successfully treated conservatively. Histological results were in accordance with preoperative diagnosis in 9 patients (81.8%). Despite the limited number of patients, our data suggest that stereotactic biopsy of brain stem lesions is a safe technique that can obtain adequate tissue for histological diagnosis, thus providing each patient with the best available treatment.