Background: To determine differences in endometrial cavity anteroposterior diameter, thickness, volume, and diameter lines of uterine body and thickness, and volume of upper, middle, and lower regions of the endometrium in infertile women using a new method for three-dimensional (3D) reconstruction based on two-dimensional (2D) ultrasound images.Methods: This retrospective cross-sectional study included a total of 81 infertile women, who underwent 2D ultrasound standard examination. We created 3D models of the uterine body, endometrial cavity, and endometrium based on 2D ultrasound images. The parameters that were measured and analyzed in a 3D plane included volume and diameter lines of endometrial cavity, surface area, thickness, volume, and diameter lines of uterine body, and surface area, thickness, and volume of upper, middle, and lower region of the endometrium. These parameters were used for comparisons between normal and arcuate uterus, between non-pregnant and pregnant infertile women, and between nulliparous and multiparous infertile women.The differences between the different regions of the endometrium and the correlations between age and the parameters were also determined in this study.