Autophagy is a homeostatic response to cellular stress. It has been shown to be potently upregulated in the heart in response to a variety of interventions. However, to date, it has not been possible to monitor autophagy without sacrificing the animal. Here we report the use of the Caliper Life Sciences Spectrum In Vivo Imaging System (IVIS) to image autophagy in homozygous transgenic mice expressing mCherryLC3 under control of the alpha myosin heavy chain promoter. Autophagy was stimulated by the administration of rapamycin (2mg/kg), and autophagosomal flux was blocked by administration of chloroquine (10mg/kg) ip. Mice were imaged at baseline and four hours later, using a protocol of 3 acquisitions of 15 seconds each. Total flux was 3.19±0.72 before drug administration and 3.93+1.10 after 4 hr, p<;0.01, n=14. These results show for the first time imaging of autophagy in hearts of live mice.