Harmonic imaging provided significant improvement in image quality by taking advantage of the scattered second harmonic (2H) component from contrast bubbles. However, differentiation between contrast and tissue (usually termed contrast-to-tissue ratio, CTR) is sometimes cumbersome and this is mainly due to tissue contamination. We have previously demonstrated, using simulations and in vitro measurements, that CTR increases as a function of the order of the harmonic number. A new contrast imaging method based on the detection of the higher harmonics was developed and termed superharmonic (SH). This technique has been shown to be more sensitive to contrast by increasing the signal from contrast and suppressing that from tissue (high CTR). The purpose of this study was to determine the clinical feasibility and usefulness of SH in patients using a commercially available contrast agent (SonoVue(R)) for quantification of myocardial perfusion. A total of 10 patients with various cardiac diseases were assessed. Apical four-chamber views were acquired using SH in triggered mode before and after contrast injection. The superharmonic was performed with a newly developed probe transmitting at 0.8 MHz with a mechanical index of 0.2. Myocardial perfusion was determined visually and analyzed quantitatively using radiofrequency (RF) processing from different regions of interest. The results showed that, before contrast injection, SH was totally blinded to tissue and no superharmonic components were generated in the image view. After administration of SonoVue(R), myocardial opacification was visualized by SH after contrast entered the myocardium. An increase of more than 15 dB in the myocardial bubbles echo compared to tissue echo was measured. In addition, the technique was used to visualize myocardial perfusion after myocardial septal ablation for hypertrophic cardiomyopathy. The clinical results showed the ability of contrast SH imaging in differentiating low and normal perfusion areas, demonstrating the high sensitivity and specificity of the technique.