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To evaluate the morphology of atrial septum by the live three-dimensional echocardi- ography (L3DE) and its value of clinical application, L3DE was performed in 62 subjects to observe the morphological characteristics and dynamic change of the overall anatomic structure of atrial sep- tum. The study examined 49 patients with atrial septal defect (ASD), including 3 patients with atrial septal aneurysm, and 10 healthy subjects. ASD in the 35 patients was surgically confirmed. The maximal diameters of ASD were measured and the percentages of area change were calculated. The parameters derived from L3DE were compared with intraoperative measurements. The results showed that L3DE could directly and clearly display the morphological features of overall anatomic structure of normal atrial septum, repaired and artificially-occluded atrial septum, atrial septal aneu- rysm. The defect area in ASD patients changed significantly during cardiac cycle, which reached a maximum at end-systole and a minimum at end-diastole, with a mean change percentage of 46.6%,ranging from 14.8% to 73.4%. The sizes obtained from L3DE bore an excellent correlation with in-traoperative findings (r=0.90). It is concluded that L3DE can clearly display the overall morphologi- cal features and dynamic change of atrial septum and measure the size of ASD area accurately, which is important in the decision to choose therapeutic protocols.