This report describes the results of CT-guided stereotactic brain biopsies performed on 50 consecutive dogs using a modified Pelorus Mark III Stereotactic System. Based on available histopathologic samples (stereotactic biopsy [n = 50], surgery [n = 17], necropsy [n = 9]) the patient population consisted of 34 dogs with primary brain tumors, 2 with invasive nasal adenocarcinomas, and 13 with non-neoplastic brain lesions. Brain tissue was not obtained from one dog. In 22 dogs a final diagnosis was made from tissue subsequently obtained from surgical resection or at necropsy. The final diagnosis was in agreement with the stereotactic biopsy diagnosis in 20 of these 22 dogs. In 17 other dogs without follow-up, stereotactic biopsy provided a diagnosis of a specific primary brain tumor subtype. Postoperative complications associated with the biopsy procedure were assessed in 41 dogs. The other 9 dogs either went directly to surgery (n = 7) or were killed (n = 2) immediately after the biopsy procedure. Thirty-six dogs recovered without apparent clinical complications. Postoperative clinical complications in the remaining 5 dogs included transient epistaxis (1 dog), transient exacerbation of cerebellar signs (1 dog), obtundation progressing to coma (1 dog), and medically uncontrollable seizures (2 dogs). The latter 3 dogs with severe neurologic complications all had large primary brain tumors and had been receiving high doses of phenobarbital and glucocorticoids to control seizures at the time of biopsy. These results suggest that this CT-guided biopsy procedure can provide an accurate pathologic diagnosis of brain lesions detected by CT and MR neuroimaging. Further refinement of both technique and case selection is expected to reduce the rate of clinical complications and to improve the accuracy of the procedure.