Mitral valve construction using extracellular matrix (ECM) is a relatively new procedure. In this case, a 15-month-old boy with a history of severe mitral valve regurgitation secondary to endocarditis underwent mitral valve surgery. Mitral valve repair was not possible, and thus, a 17 mm extracellular matrix cylinder valve (ECM-CV) was constructed for valve replacement. The ECM-CV is clearly imaged using echocardiography, especially three-dimensional imaging, that helped define valve function. As the use of ECM for valve construction increases, echocardiography will play an essential role in evaluating the function and mechanics of these novel valves.