The Wohl Institute for Translational Medicine 1 article published in JoVE Biology Investigating Cardiac Metabolism in the Isolated Perfused Mouse Heart with Hyperpolarized [1-13C]Pyruvate and 13C/31P NMR Spectroscopy David Shaul1,2, Gal Sapir1,2, Naama Lev-Cohain1, Jacob Sosna1, J. Moshe Gomori1, Rachel Katz-Brull1,2 1Department of Radiology, Hadassah Medical Organization and Faculty of Medicine, Hebrew University of Jerusalem, 2The Wohl Institute for Translational Medicine We describe an experimental setup for administrating hyperpolarized 13C-labeled metabolites in continuous perfusion mode to an isolated perfused mouse heart. A dedicated 13C-NMR acquisition approach enabled the quantification of metabolic enzyme activity in real-time, and a multiparametric 31P-NMR analysis enabled the determination of the tissue ATP content and pH.