Understanding the rate at which specific skills are acquired and the rate at which they decay is critical for designing training curricula, simulation-based training, certification standards, and refresher training. Retention rates of specialized medical skills are of particular interest and relevance to the military due to the nature of military deployment cycles. For example, surgical skills such as those required for performance of laparoscopic surgical procedures have been reported to decay during long military deployments as these specialized skills are not utilized within deployed settings. In an effort to better understand the nature of medical skills acquisition and decay, a study was conducted examining initial training and retention over several weeks of standardized laparoscopic surgical psychomotor skills using the Fundamentals of Laparoscopic Surgery (FLS) manual skills training platform. Of particular interest in this study was the role of skill acquisition and retention with the dominant versus the non-dominant hand. Expert surgeons have indicated that ambidexterity plays a significant role in surgical skill proficiency. The results of this study indicate significant differences in performance between the dominant and non-dominant hands during the early stages of training, with ambidexterity increasing as trainees reach proficiency. This research lays the groundwork for a longitudinal research study in which retention of the trained skills will be assessed following a 6-month period of nonuse. Implications for objective assessment of medical skill acquisition, proficiency, and retention are discussed, including implications for training and retention of a broad range of medical skills involving psychomotor components using simulation-based training.