Abstract This study was carried out to evaluate a method used to measure three-dimensional (3D) cognitive fatigue based on the pupillary response. This technique was designed to overcome measurement burdens by using non-contact methods. The pupillary response is related to cognitive function by a neural pathway and may be an indicator of 3D cognitive fatigue. Twenty-six undergraduate students (including 14 women) watched both 2D and 3D versions of a video for 70 min.