Abstract: Objective To investigate the feasibility of tracking bone mesenchymal stem cell (BMSCs) dual-labeled with polyethylenimine 2k-superparamagnetic iron oxide (PEI2k-SPIO) and Luciferase transplantation for acute myocardial infarction in vivo by using magnetic resonance imaging (MRI) and fluorescence imaging. Methods BMSCs/Luciferase was incubated with culture medium containing PEI2k-SPIO for 24 h. Prussian-blue staining and MTT were used to assess the efficacy and safety of labeling with PEI2k-SPIO. Guided with echocardiography, the dual-labeled BMSCs were injected into the margin of infarction myocardium. MRI and fluorescence imaging were performed to monitor the cells in vivo at different times (1,2,3,7 d). Results As demonstrated by MTT, there was no significant difference in survival rate between the labeled and unlabeled cells (P>0.05). Within a week after transplantation, all PEI2k-SPIO-labeled BMSCs showed a significant decreased signal on MRI. Dual-labeled BMSCs were detected bioluminescence with fluorescence imaging, but disappeared after one week. Conclusion Multi-modality imaging can not only trace the location of labeled BMSCs but also demonstrate the survival of labeled BMSCs in vivo.