Objective To investigate the clinical value of method of detecting sentinel lymph node (SLN) by different frozen sections in the surgical treatment of breast cancer. Methods Sixty-seven breast cancer patients were given methylene blue and successfully underwent sentinel lymph node biopsy. Intraoperative frozen sections of SLN were made and HE staining was performed. The pathological results of single slice, 3 slice and 6 slice were recorded. Postoperative routine HE staining of SLN paraffin sections was performed and considered as the standard of the diagnosis of SLN metastasis. Results of pathological diagnosis of different frozen sections were compared with the results of paraffin sections. Results The routine paraffin section results of the 67 patients revealed positive SLN in 19 cases, false negative SLN in 3 cases, the false negative rate was 13.6% (3/22). Comparison of the results of different slicing methods of frozen sections with paraffin sections of SLN showed the sensitivity of single slice was 21.1% (4/19), the coincidence rate was 73.1% (49/67), the specificity was 100% (45/45); the sensitivity of 3 slice was 31.6% (6/19), the coincidence rate was 76.1% (51/67), the specificity was 97.8% (44/45); the sensitivity of 6 slice was 84.2% (16/19), the coincidence rate was 91.1% (61/67), the specificity was 95.6% (43/45). The sensitivity and coincidence rates of the 6 slice were significantly higher than those of 3 slice and single slice, the differences were significant (P < 0.05). The difference between the single slice group and the 3 slice group were not significant (P > 0.05). Conclusions SLN intraoperative frozen pathological diagnosis is a reliable and fast method for diagnosis of SLN metastais. Multilayer section method can improve the detection rate of SLN metastasis, the results of 6 layer frozen section is close to postoperative pathological results of paraffin section.