Abstract: Non-invasive quantitative analysis of the heart walls thickness is a fundamental step in diagnosis and discrimination of heart disease. Thickness measurements in 2D echocardiographic images have many applications in research and clinic for assessing of wall stress, wall thickening and viability parameters. Regarding to interventricular septum wall thickness measurement by conventional manual method is more dependent on sonographer experiment; this encouraged these researchers to develop a semi-automatic computer algorithm in accessing to interventricular septum segments thickness. We proposed and carried out a computerized algorithm for wall thickness measurements in 2D echocardiographic image frames. In this program, wall thickness measurement is based of intensity profile function and adaptive bilateral thresholding operation. For validation, thicknesses of septum base and mid segments were estimated in constituent image frames with use of proposed method and then were compared with conventional manual results at same images of the cardiac cycle by statistical methods. In our sample image frames (240 corresponding segments; with different rang of image quality), a bias of 0.10 and 0.12 mm with SD differences of ± 0.81 and ± 0.72 mm and correlation coefficients of 0.87 and 0.89 were found in base and mid segments, respectively. Interobserver variability using the Computer-Assisted Method (CAM) and Conventional Manual Method (CMM) were 4.0 and 4.7% for the basal and 2.8 and 3.9% for the middle segments. The method introduced in the present study permits precise thickness assessment of base and mid segments of the interventricular septum wall and has high concordance with CMM.