To evaluate the feasibility of real-time myocardial contrast echocardiography (RTMCE) by quantitative analysis of myocardial perfusion in rabbits, transthoracic RTMCE was performed in 10 healthy rabbits by using continuous infusion of SonoVue into the auricular vein. The short axis view at the papillary muscle level was obtained. The duration of the time that the contrast took to appear in right heart, left heart and myocardium was recorded. The regional myocardial signal intensity (SI) versus refilling time plots were fitted to an exponential function: y(t) =A(1-e(-beta(t-t0))) + C, where y is SI at any given time, A is the SI plateau that reflects myocardial blood volume, and beta is the slope of the refilling curve that reflects myocardial microbubble velocity. The A, beta and Axbeta values at different infusion rate of SonoVue were analyzed and the A, beta and Axbeta values in each segment in the short axis view at the papillary muscle level were compared. All the animal experiments were successful and high-quality images were obtained. The best intravenous infusion rate for SonoVue was 30 mL/h. The contrast appeared in right heart, left heart and myocardium at 7.5+/-2.2 s, 9.1+/-2.4 s and 12.2+/-1.6 s respectively. After 16.6+/-2.3s, myocardial opacification reached a steady state. The mean A, beta and Axbeta value in the short axis view at the papillary muscle level were 9.8+/-3.0 dB, 1.4+/-0.5 s(-1) and 13.5+/-3.6 dBxs(-1) respectively. A, beta and Axbeta values showed no significant differences among 6 segments. It was suggested that RTMCE was feasible for quantitative analysis of myocardial perfusion in rabbits. It provides a non-invasive method to evaluate the myocardial perfusion in rabbit disease models.