Background: Acute myocardial infarction (MI) is the primary factor leading to cardiovascular diseases, which are the main causes of morbidity and mortality in developed countries. Mesenchymal stem cell (MSC)-derived exosomes have been reported to improve heart function after MI; however, the molecular mechanisms responsible for this are unknown. In vivo imaging can reveal the trafficking process and in vivo biodistribution of exosomes, which may provide an insight into the communication mechanisms and pharmacokinetics of exosomes.Methods: Glucose modified gold nanoparticles were used to label MSC-derived exosomes, aimed at minimizing membrane damage and maintaining the integrity of the exosomes. After labeling, the exosomes were visualized by in vivo computed tomography (CT) imaging to determine the biodistribution at 4 and 24 h after injection into a MI mouse model.Results: MSC-derived exosomes were successfully labeled by glucose modified gold nanoparticles and CT imaging of these labeled exosomes indicated that MSC-Exo remained in the MI area for up to 24 h after intramyocardial injection. Additionally, few MSC-Exo were observed in some other organs, particularly the liver, spleen, and kidney.Conclusions: A gentle method was used for loading GNPs into exosomes, and their successful labeling without causing aggregation was verified. In vivo CT imaging revealed the retention of MSC-Exo in the MI area, indicating their usefulness for improving heart function after infarction.