Thyroid and parathyroid glands play a vital role in regulating the body's metabolism and calcium levels. Surgical removal of the glands is the main treatment for both thyroid cancer and parathyroid adenoma. In thyroidectomy and parathyroidectomy, it's very important to differentiate thyroid, parathyroid, and the other tissues around the neck. Traditionally, physicians use ultrasound guided fine needle aspiration (FNA) to evaluate thyroid nodules, but up to 30% of FNA results are “inconclusive”. The sestamibi scan can localize parathyroid adenoma, but currently it only has 50% accuracy. Here we applied the emerging CARS technique to image both thyroid and parathyroid tissues, which has potential to be used in real-time in vivo examination of different structures. We also developed algorithms to differentiate different cellular structures based on CARS images. When incorporated with a fiber optic endoscope in the future, CARS imaging technique can help surgeons identify cancerous thyroid tissue intraoperatively, preserve good parathyroid glands during thyroidectomy and find parathyroid adenoma during parathyroidectomy.