Introduction. This research investigated differences in the effects of a state-of-art stereoscopic 3-dimensional (3D) display and a traditional 2-dimensional (2D) display in simulated laparoscopic surgery over a longer duration than in previous publications and studied the learning effects of the 2 display systems on novices. Methods. A randomized experiment with 2 factors, image dimensions and image sequence, was conducted to investigate differences in the mean movement time, the mean error frequency, NASA-TLX cognitive workload, and visual fatigue in pegboard and circle-tracing tasks. Results. The stereoscopic 3D display had advantages in mean movement time (P < .001 and P = .002) and mean error frequency (P = .010 and P = .008) in both the tasks. There were no significant differences in the objective visual fatigue (P = .729 and P = .422) and in the NASA-TLX (P = .605 and P = .937) cognitive workload between the 3D and the 2D displays on both the tasks. For the learning effect, participants who used the stereoscopic 3D display first had shorter mean movement time in the 2D display environment on both the pegboard (P = .011) and the circle-tracing (P = .017) tasks. Conclusions. The results of this research suggest that a stereoscopic system would not result in higher objective visual fatigue and cognitive workload than a 2D system, and it might reduce the performance time and increase the precision of surgical operations. In addition, learning efficiency of the stereoscopic system on the novices in this study demonstrated its value for training and education in laparoscopic surgery.