In vivo assessment of cardiac structure and function in small animals is an important experimental goal, but currently available techniques have significant limitations. A commercially available intravascular ultrasound (IVUS) system was adapted to perform transesophageal echocardiography (TEE) in rats. Twelve Sprague-Dawley rats (270-370 g) were anesthetized with intraperitoneal pentobarbital sodium. A 4.3-Fr, 30-MHz or an 8-Fr, 20-MHz IVUS catheter was inserted into the esophagus to obtain long-axis views of the aortic arch, short-axis views of the ascending aorta, and long-axis views of the pulmonary artery. A preshaped, 8-Fr, 20-MHz catheter was used to obtain short-axis images of the left ventricle (LV) at the midpapillary muscle level, which were used to measure LV diastolic and systolic dimensions (diameters) and to calculate LV mass and fractional shortening. Measurements by TEE were compared with those obtained by transthoracic echocardiography in 6 of 12 rats. Postmortem, the LV was weighed to determine actual LV mass. The correlation coefficients between TEE- and transthoracic echocardiography-calculated LV mass and actual LV mass were 0.94 and 0.88, respectively, and had a good agreement with actual LV mass. Inter- and intraobserver variability of TEE measurements was <10%. IVUS instrumentation may offer an alternative technique for the accurate, serial assessment of LV dimensions, mass, and systolic function and a means of imaging the great vessels in small laboratory animals.