The identification and preservation of parathyroid glands (PGs) is a major issue in thyroidectomy. The PG is particularly difficult to distinguish from the surrounding tissues. Accidental damage or removal of the PG may result in temporary or permanent postoperative hypoparathyroidism and hypocalcemia. In this study, a novel method for identification of the PG was proposed based on laser-induced breakdown spectroscopy (LIBS) for the first time. LIBS spectra were collected from the smear samples of PG and non-parathyroid gland (NPG) tissues (thyroid and neck lymph node) of rabbits. The emission lines (related to K, Na, Ca, N, O, CN, C-2, etc.) observed in LIES spectra were ranked and selected based on the important weight calculated by random forest (RF). Three machine learning algorithms were used as classifiers to distinguish PGs from NPGs. The artificial neural network classifier provided the best classification performance. The results demonstrated that LIBS can be adopted to discriminate between smear samples of PG and NPG, and it has a potential in intra-operative identification of PGs. (C) 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement