The recently developed method of ultrarapid immunohistochemistry (IHC) was applied to the intraoperative examination of sentinel lymph nodes (SLNs) in breast cancer patients. In a prospective study of 50 patients with invasive breast carcinomas, a total of 60 SLNs were studied. Among them, 33 SLNs from 30 patients were studied intraoperatively using a direct immunoperoxidase method with anticytokeratin antibody clone MNF116. This technique has a turnaround time of less than 20 minutes. Ultrarapid IHC revealed 15 positive SLNs compared to 14 positive SLNs using hematoxylin and eosin (H and E) frozen sections. The one SLN missed in H and E frozen sections presented with cytokeratin-positive isolated tumor cells in the lymph node sinus. After paraffin embedding, H and E-stained serial step sections of the SLN specimens detected another two patients with isolated tumor cells. We also examined the remaining axillary lymph nodes (ALNs) by H and E-stained serial step paraffin sections. From 17 of the 30 patients with positive SLNs, 6 patients also had metastatic involvement of the ALNs of level I or II. Thus ultrarapid IHC was a very sensitive and rapid technique for the intraoperative detection of metastatic involvement of SLNs in breast cancer patients. This technique may be a useful complementary tool for the intraoperative study of SLNs, particularly in tumors that are a diagnostic challenge, such as lobular carcinoma.