Intraoperative pathologic diagnosis for central nervous system (CNS) tumors is important to determine the neurosurgery procedure. But sometimes the differential diagnosis between glioma and lymphoma, or glioma and metastatic tumors is difficult for a pathologist during a short time, especially when the specimen is small or the frozen section has ice crystals. Immunohistochemistry (IHC) is a very useful method for diagnosis, but the traditional immunohistochemical method is time-consuming and not suitable intraoperatively. In this study, we chose Cytokeratin-pan, GFAP, and LCA as three immunohistochemical indicators. Intraoperative IHC was done by Novodiax ihcDirect technology combined with Leica Bond auto-staining. Compared with the manual method recommended for the reagents (Novodiax ihcDirect), the results show that auto-staining has better stability and high reproducibility in coloration, which has broad prospects for future application.