It is difficult for conventional transthoracic echocardiography (TTE), by which precise and accurate images of interatrial septum (IAS) can not be acquired, to diagnose patent foramen ovale (PFO) clearly. To evaluate the diagnostic value of biplanar transesophageal echocardiography (TEE) for PFO, TTE and biplanar TEE were performed simultaneously in 270 patients. It was found that in 7 patients patent foramen ovale was detected only through longitudinal planes of biplanar TEE. IAS, which consists of primitive septum and membrane of fossa ovalis, can be directly visualized by two-dimensional images of TEE; in patients with PFO, a dull color flow, which shunts from the right atria to the left atria through the gap between primitive septum and fossa ovalis, can be detected by color Doppler flow images. Furthermore, some right-to-left shunting microbubbles through the valve of patent fossa ovalis can be discovered by cardiac acoustic contrast echocardiography. In conclusion, biplanar TEE combined with color Doppler image and cardiac acoustic contrast facilitates a definite diagnosis of patent foramen ovale as the excellent anatomic images of IAS can be obtained from multiple views under this kind of performance.