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Fluorescence imaging can track the expression of fluorescent protein and 18F-fluorodeoxyglucose based positron emission tomography ( [18F]FDG-PET) can evaluate the changes of [18F]FDG uptake in tumor cells during the antitumor treatment. In this work, fluorescence imaging and [18F]FDG-PET were both employed to monitor tumor response to Au nanorod-indocyanine green (AuNR-ICG) conjugates mediated therapy in a subcutaneous MDA-MB-231 mouse xenograft model. Serial fluorescence and [18F]FDG-PET images following the antitumor treatment were obtained and quantitative analysis revealed significant decreases in fluorescence intensity and metabolic activity in tumors treated with AuNR-ICG conjugates under near infrared laser irradiation. The results suggest that the combination of fluorescence and [18F]FDG-PET imaging can provide a noninvasive tool to assess the tumor response to antitumor therapy on a molecular scale.