Abstract: Background　Rapid and accurate identification of resected tissues is crucial during breast cancer surgery. The Sparameter- based method for distinguishing biological tissues shows potential, but the classical open-ended coaxial probe method requires multiple calibrations to utilize the S11 parameter for identification, limiting its clinical application. The novel transmitreceive integrated probe method eliminates the need for complex calibration and can simultaneously acquire both S11 and S21 parameters, offering a new approach for rapid biological tissue detection.Objective　To validate the potential application of the novel transmit-receive integrated probe in the rapid identification of biological tissues. Methods　Using tumor-bearing mice as subjects, S-parameters of skin, fat, muscle, and tumor tissues were measured using both the open-ended coaxial probe and the novel transmit-receive integrated probe. The tissue discrimination capability was evaluated through magnitude difference analysis, multifrequency point (5, 10, 15, and 20 GHz) statistical testing, and machine learning algorithms. Furthermore, the clinical applicability of this method was verified using surgically resected specimens (including tumor, gland, fat, skin, and adjacent normal tissues). Results　Measurements on tumor-bearing mice using the open-ended coaxial probe showed no significant difference in S11 parameters between tumor and fat tissues at 5 GHz, nor between tumor and muscle tissues at 10, 15, and 20 GHz. In contrast, the novel transmit-receive integrated probe detected significant differences (P＜0.001) in both S11 and S21 parameters between tumor and the three types of normal tissues at all four frequency points. Machine learning analysis results from both tumor-bearing mice and surgically resected specimens indicated that combining S11 and S21 parameters improved classification performance, with the S21 parameter significantly enhancing the ability to identify normal or adjacent normal tissues. Conclusion　This study systematically validates the effectiveness of the novel transmit-receive integrated probe for rapid discrimination of biological tissues through experiments on tumor-bearing mice and surgically resected specimens, preliminarily demonstrating its clinical applicability and providing new technical support for real-time tissue identification during breast cancer surgery.