The development of experimental animal models for head and neck tumors generally rely on the bioluminescence imaging to achieve the dynamic monitoring of the tumor growth and metastasis due to the complicated anatomical structures. Since the bioluminescence imaging is largely affected by the intracellular luciferase expression level and external D-luciferin concentrations, its imaging accuracy requires further confirmation. Here, a new triple fusion reporter gene, which consists of a herpes simplex virus type 1 thymidine kinase (TK) gene for radioactive imaging, a far-red fluorescent protein (mLumin) gene for fluorescent imaging, and a firefly luciferase gene for bioluminescence imaging, was introduced for in vivo observation of the head and neck tumors through multi-modality imaging. Results show that fluorescence and bioluminescence signals from mLumin and luciferase, respectively, were clearly observed in tumor cells, and TK could activate suicide pathway of the cells in the presence of nucleotide analog-ganciclovir (GCV), demonstrating the effectiveness of individual functions of each gene. Moreover, subcutaneous and metastasis animal models for head and neck tumors using the fusion reporter gene-expressing cell lines were established, allowing multi-modality imaging in vivo. Together, the established tumor models of head and neck cancer based on the newly developed triple fusion reporter gene are ideal for monitoring tumor growth, assessing the drug therapeutic efficacy and verifying the effectiveness of new treatments.