Abstract The heart requires a constant supply of energy to sustain contractile function, which is supplied by hydrolysis of adenosine triphosphate derived primarily from the metabolism of fatty acids and carbohydrates. Understanding how production of adenosine triphosphate is regulated in the heart is critical to an understanding of how alterations in energy metabolism contribute to the severity of cardiac disease. A number of techniques can be used to measure energy metabolism in the heart. They include biochemical measurement of metabolites and enzymes of intermediary metabolism, measurement of arteriovenous differences in carbon substrate extraction by the heart, measurement of high-energy phosphates with 31P nuclear magnetic resonance, measurement of the rate of flux through the pathways of intermediary metabolism with 14C- and 3H-labeled carbon substrates, measurement of tricarboxylic acid cycle activity with 13C nuclear magnetic resonance, and measurement of glucose uptake and oxidative metabolism with positron emission tomography. Each of these techniques has advantages and limitations.