Despite the potential ability of left ventricular (LV) shape analysis to provide independent information complementary to ventricular size and function, in clinical practice only ejection fraction (EF) is currently assessed while LV shape is not routinely quantified. Moreover, geometric assumptions in the computation of EF from multiple two-dimensional (2-D) cut-planes by disc summation or area-length methods, introduce inaccuracies in the estimates. Also, previous approaches for the quantification of LV shape were based on geometric modeling and, as a result, proved inaccurate. Our aims were (1) to develop and test a three-dimensional (3-D) technique for direct quantification of LV shape from real-time 3-D echocardiographic (RT3DE) images without the need for geometric modeling using a new class of LV shape indices; and (2) to study the relationship between these indices and ventricular size and function in normal and abnormal ventricles. Spherical (S), ellipsoidal (E) and conical (C) shape indices were calculated using custom software for analysis of transthoracic RT3DE images on both global and regional levels and initially tested on computer simulated objects of different shapes. The feasibility of using these indices to differentiate between normal and abnormal ventricles was tested in three groups of patients: normal volunteers (NL, n=9), dilated cardiomyopathy (DCM, n=9) and coronary artery disease with apical regional wall motion abnormalities (RWMA, n=9). Computer simulation demonstrated that these shape indices are size-independent and can correctly classify the simulated objects. In human ventricles, both S and C but not E correlated well with LV volumes and EF. Also, S and C changed throughout the cardiac cycle while E remained almost constant. In addition, both regional and global S and C were able to identify differences between NL and abnormal ventricles: normal ventricles were less spherical and more conical than those of patients with DCM at both end-systole and end-diastole (p<0.05) both globally and regionally. In contrast, in patients with RWMA, similar differences were noted only at end-systole, both on a global level and in the apical region. In this study, we demonstrated the feasibility of quantifying LV shape from transthoracic RT3DE images at both global and regional levels. Potentially, such 3-D shape analysis could be combined with conventional evaluation of LV volume and function to provide a more comprehensive assessment of left ventricular performance.