Objective The goal here was to conduct a preliminary evaluation of the efficacy of a conjugate (NIRG) of heptamethine carbocyanine dye (IR-783) and the chemotherapeutic drug gemcitabine for tumor fluorescent imaging and targeted therapy. Methods First, the liver cancer cell line Hep3B was used to verify the specific absorption of NIRG, and nude mice as models of tumors from a subcutaneous xenograft of Hep3B cells were prepared to observe the effect of the NIRG fluorescence imaging in this CDX model. Three subcutaneous PDX models of liver cancer cases (C64003, C34566 and B66873) were prepared to study the effect of NIRG tumor-targeted fluorescence imaging and targeted therapy. Simultaneously, the expression of the hepatoma-specific antigen AFP was observed by immunohistochemistry in tumor tissues of these models. In addition, the expression of the antigens HIF1α and OATP associated with the specific uptake of NIRF dye was observed. Results The results showed that the binding sites of NIRG in liver cancer cell line Hep3B cells were mitochondria and lysosomes. NIRG-specific tumor sites were found in the subcutaneous CDX model of liver cancer, and the NIRF signal was enhanced with the growth of the tumor. NIRF signal and the luminescence signal had a good correlation (R² =0. 99716). The IHC results showed that AFP was highly expressed in the three PDX models, and HIF1α and OATP were also highly expressed. The fluorescence imaging results showed that NIRG had good tumor targeting and could inhibit tumor growth, showing a good therapeutic effect. Conclusions The conjugate NIRG has characteristics suitable for both fluorescence imaging and tumor targeting to deliver chemotherapeutic drugs. It is potentially an effective novel imaging and targeting agent for treatment of human liver cancer.