We sought to summarize the published evidence regarding the measurement of infarct size by serum markers, technetium-99m sestamibi single-photon emission computed tomography (SPECT) myocardial perfusion imaging, and magnetic resonance imaging. The measurement of infarct size is an attractive surrogate end point for the early assessment of new therapies for acute myocardial infarction. For each of these three approaches, we reviewed reports published in English providing the clinical validation for the measurement of infarct size and the relevant clinical trial experience. The measurement of infarct size by serum markers has multiple theoretical and practical limitations. The measurement of troponin is promising, but the available data validating this marker are limited. Sestamibi SPECT imaging has five separate lines of published evidence supporting its validity and has received extensive study in multicenter trials. Magnetic resonance imaging has great promise but has less clinical validation and no multicenter trial experience. Therefore, SPECT sestamibi imaging is currently the best available technique for the quantitation of infarct size to assess the incremental treatment benefit of new therapies in multicenter trials of acute myocardial infarction.