The use of perfusion imaging in the acute phase of myocardial infarction has been facilitated by the introduction of technetium 99m-labeled sestamibi (99mTc-sestamibi). Because of minimal redistribution, myocardium at risk can be quantified without delaying reperfusion therapy. The use of perfusion imaging with 99mTc-sestamibi has been extensively validated in a series of important animal studies in contrast to other methods used to assess outcome from acute myocardial infarction. This has important implications regarding the assessment of reperfusion therapy. With an accurate means to define myocardium at risk, myocardial salvage can be measured for specific therapies or patient subsets. Such measures also have clinical utility for the care of individual patients. Infarct size measures with 99mTc-sestamibi are accurate and predictive of subsequent left ventricular remodeling as well as prognosis. The identification of jeopardized myocardium in patients with nondiagnostic electrocardiograms and the noninvasive prospective measurement of collateral blood flow before reperfusion therapy are two new areas where perfusion imaging has special clinical use. Because of the ability of perfusion imaging with 99mTc-sestamibi to measure most of the variables known to determine infarct size, comparative clinical trials can be accomplished by using relatively small sample sizes. This has important implications regarding the assessment of new therapies for acute myocardial infarction.