To determine the accuracy of intraoperative cytopathological diagnosis compared with conventional histopathological diagnosis, the authors obtained 100 specimens from masses of various organ systems chosen randomly from 65 dogs, 30 cats, and five exotic animals. Of the 100 specimens, a specific diagnosis was obtained in 42%, the correct pathological process (i.e., mesenchymal neoplasia, epithelial neoplasia, round cell neoplasia, or inflammation) was identified in 41%, in 1% the diagnosis was deferred, and in 16% an incorrect diagnosis was obtained. The overall accuracy rate of intraoperative cytopathological examination was 83%, which increased to 90% by the exclusion of splenic masses. The accuracy rate of diagnosing neoplasia was 87%, with a sensitivity of 89% and a specificity of 100%. Intraoperative cytopathological examination is an accurate diagnostic method with good sensitivity and specificity for the identification of neoplasia.