In this study, the impact of two-dimensional (2D) and three-dimensional (3D) vision on laparoscopic performance of novice surgeons is examined. Twenty-five novice surgeons were directed to complete four basic tasks from European Training in Basic Laparoscopic Urological Skills (E-BLUS) with both 2D and 3D systems in a random order: task 1: needle guidance, task 2: cutting a circle, task 3: laparoscopic suturing, and task 4: pegs transfer. Quality and quantity scores for each task were measured. Participants completed all of the tasks in one modality of vision and than switched to the other. NASA Task Load Index was used for subjective workload assessment. Statistically significant differences in favor of 3D vision were detected in tasks 1 and 4 both in terms of quality and quantity. In task 2 and task 3, a significantly better performance was observed with the 3D vision only in quantity assessment. The participants who started the tasks in the 3D vision were better in performing the skills in 2D when compared to the participants who started with 2D vision. Overall, the participants reported a better perception of depth and spatial orientation with the 3D mode. Subjective work load was also lower for the tasks performed in 3D. Novice surgeons tended to perform better and felt much more comfortable with 3D in comparison to 2D laparoscopy. Even though previous task experience seemed to have an important impact on laparoscopic performance regardless of imaging modality, 3D laparoscopy seemed to facilitate the learning for novice surgeons.