This chapter investigates one of the advanced and latest imaging techniques, near-infrared fluorescence (NIRF) imaging in head and neck cancer surgery. NIRF technology enhances tumor margin assessment, lymphatic metastasis imaging, and ex vivo specimen analysis. It employs both nonspecific target dyes and specific target probes to accurately delineate cancerous tissues and effectively identify the metastatic status of lymph nodes. The analysis of ex vivo specimens facilitates the evaluation of positive status of tumor margins. Moreover, NIRF technology also plays a crucial role in the treatment strategies for nerve protection, parathyroid gland preservation, and thoracic duct safeguarding. NIRF imaging is increasingly being integrated into standard head and neck tumor surguries, potentially leading to significant improvements in surgical outcomes and patient prognosis.