Assessment of visual fatigue caused by stereoscopic disparity based on multimodal measurement

•Subjective and objective measurements showed that visual fatigue was affected by stereoscopic horizontal disparity.•Subjective questionnaire, ophthalmological parameters, eye movement and electrocardiogram (ECG) metrics can distinguish visual fatigue caused by 3D display.•Subjects felt more headache, uncomfortable and dry eyes after watching 70-minute enhanced 3D display with larger negative disparity than normal 3D display.•Visual function tear film break-up time (TBUT) reduced, mean microsaccade pupil diameter (MMPD) increased and heart rate variability (HRV) changed more significantly after watching enhanced 3D display.•Enhanced 3D display induced more visual fatigue significantly than normal 3D display.