Balloon occlusion and release during elective percutaneous coronary intervention (PCI) provides a unique opportunity to study dynamic temporal alterations in myocardial perfusion in a controlled setting. These changes in flow and volume mimic those that occur during presentation with, and successful therapy of, ST-segment elevation acute myocardial infarction (AMI). Eleven patients underwent myocardial contrast echocardiography (MCE) using a continuous infusion of Definity at baseline, during coronary occlusion, and during reactive hyperemia immediately after balloon deflation. Fifty separate flow state sequences were acquired, and off-line analysis was performed to determine myocardial contrast intensity within a region of interest in the distribution of the left anterior descending artery (LAD). A reader blinded to flow state also performed qualitative evaluation (perfusion or lack of perfusion). Quantitative analysis demonstrated significant differences in myocardial contrast intensity by flow state (p = 0.0001 for occlusion vs reperfusion). Qualitative assessment demonstrated a high rate of correct classification (92%). Real-time myocardial perfusion assessment using MCE accurately differentiates coronary occlusion and reactive hyperemia in humans by qualitative and quantitative assessment. This technique may be clinically useful in assessing the efficacy of thrombolytic therapy in ST-segment elevation AMI and in clinical trial assessment of new drugs and devices aimed at limitation of infarct size.