Objective：To study sentinel lymph node biopsy（SLNB） in breast cancer patients by using 1% isosulfan blue or methylene blue. Method：206 patients were randomly divided into 2 groups，IB and MB，with injection different dye through peri-nipple and breast parenchyma. All patients undergoing modified radical mastectomy or radical mastectomy with axillary lymph nodes dissection（ALND）, and then the nodes in the ALND specimen were processed for histological examination. 8 female adult rabbits were randomly divided into 2 groups to study the dying time，number and fading time of the sentinel lymph nodes with different dyes. Results:In isosulfan blue group，SLNs were identified in 94 out of 102 patients，with a detection rate of 92.16%. The sensitivity of SLNB in this study was 92.31%，accuracy was 90.91%，false negative rate was 6.06%，false positive rate was 0.In methylene blue group，SLNs were identified in 97 out of 104 cases，with a detection rate of 93.27%. The sensitivity of SLNB in this study was 93.75%，accuracy was 91.35%, false negative rate was 6.25%，false positive rate was 0. No statistical significance was shown between isosulfan blue group and methylene blue group（P > 0.05）. In the rabbits group，the average staining time of the IB and MB group was 8.75s and 13.13s respectively. There are significant difference of the dying time，number and fading time of the sentinel lymph nodes between two groups（P = 0.000）. Conclusion：SLNB using methylene blue possesses the same characteristics of safety and feasibility as using isosulfan blue. Methylene blue is very cheap and can be widely used in the clinic.