Objective The aim of this study was to access method by which identify viable myocardium by two-dimensional strain analysis combined with PLGA contrast enhanced low dose dobutamine stress echocardiography. Methods Thirty cute ischemia reperfusion canine models were reproduced successfully. While real-time myocardial contrast (enhanced with PLGA contrast agents) performed, images were acquired and saved at baseline, rest , and the period low dose dobutamine administration. Measurements of regional peak-systolic strain in longitudinal, radial and circumferential motion on all segments were performed respectively under different condition. Results Compared with nonviable myocardial segment, LS, RS and CS of viable segment increased significantly during dobutamine stress (p <0.05; p <0.01), and LS, RS and CS of the former did not present significant variance at same time (p >0.05). Respective sensitivities for LS, RS, and CS were 81.03%, 77.59%, and 72.41%; respective specificities were 81.3%, 84%, and 78%. Conclusion Two-dimensional strain analysis combined with low dose dobutamine stress echocardiography enhanced with PLGA contrast agent can afford a novel way to distinguish viable myocardium from the non-viable myocardium with a incremental accuracy.