AbstractAutostereoscopic displays are likely to become widely used products in the future. However, certain physiological factors, especially visual comfort, limit their development. In this study, four observational parameters – ambient illuminance, image content, scaling ratio, and horizontal distance between major and minor objects – were evaluated to determine the degree of visual comfort offered by 3D computer‐generated images on an autostereoscopic display. Visual comfort score with the range of 0–1 is designed to represent the degree of visual comfort for the 3D images with different manipulations of ambient illuminance, image content, scaling ratio, and horizontal distance between major and minor objects in this study. Subjects were asked to indicate images that produced discomfort. The proportion of images for each condition where participants indicated that viewing the image was comfortable was computed. Images receiving a proportion of 0.5 or greater were classified as acceptable. The disparity ranges over which acceptable images were attained for each participant and for each condition were analyzed with analysis of variance. The analytical results indicate that ambient illuminance and image content have a significant effect on the acceptable disparity range, while scaling ratio and horizontal distance between major and minor objects did not.