In single-access surgery, instruments enter the abdominal cavity through only 1 incision, the position of the instruments relative to each other is different compared with that in conventional laparoscopy. Changes in instrument configuration may increase task complexity and therefore affect tissue handling skills. The aim of this study is to determine if a relation exists between instrument configuration and tissue interaction force. A study was performed to investigate the differences in manipulation force between a single-port (SP) and 2-port (TP) instrument configuration in a standard box trainer. A force platform was placed under a tissue manipulation task in a box trainer and used to measure the pulling forces and trial time. A total of 28 medical students with no previous experience in laparoscopic surgery were divided into 2 equal groups. Group 1 trained the task 6 times with the TP configuration and subsequently performed 6 trials with the SP configuration. Group 2 used the configurations in the opposite order. For both groups, the learning curves of the maximum force and task time were compared. Time and maximum pulling forces were significantly different between the 2 instrument configurations. In both groups, the participants used significantly more force in the SP configuration than in the TP configuration. The force data indicate that the increased complexity in instrument handling with straight instruments in a SP configuration increases the tissue manipulation force. Furthermore, the tissue handling skills of novices who mastered the task with the TP configuration decreased after switching to the SP configuration.