A noninvasive, whole-body, real-time fluorescence optical imaging of stable high-level green fluorescent protein (GFP)-expressing human adenoid cystic carcinoma (ACC-M-GFP) was demonstrated for in vivo visualization of metastatic behavior in nude mice. Five-week-old female nude mice were injected with ACC-M-GFP in the primary organ: submandibular gland. Metastases were only visualized by GFP expression in the lung. However, metastatic lesions of ACC-M-GFP in the lung, muscle, bladder and bony were found by imaging of GFP expression in intact mice through tail vein injection of ACC-M-GFP cells. The construction of highly fluorescent and stable GFP transfectants of ACC-M has revealed the multi-organ metastatic capability of ACC-M cells through this optical imaging.