The prefrontal cortex (PFC) is known to be vital for acquisition of visuomotor skills, but its role in the attainment of complex technical skills which comprise both perceptual and motor components, such as those associated with surgery, remains poorly understood. We hypothesized that the prefrontal response to a surgical knot-tying task would be highly dependent on technical expertise, and that activation would wane in the context of learning success following extended practice. The present series of experiments investigated this issue, using functional Near Infrared Spectroscopy (fNIRS) and dexterity analysis to compare the PFC responses and technical skill of expert and novice surgeons performing a surgical knot-tying task in a block design experiment. Applying a data-embedding technique known as Isomap and Earth Mover's Distance (EMD) analysis, marked differences in cortical hemodynamic responses between expert and novice surgeons have been found. To determine whether refinement in technical skill was associated with reduced PFC demands, a second experiment assessed the impact of pre- and post-training on the PFC responses in novices. Significant improvements (p < 0.01) were observed in all performance parameters following training. Smaller EMD distances were observed between expert surgeons and novices following training, suggesting an evolving pattern of cortical responses. A random effect model demonstrated a statistically significant decrease in relative changes of total hemoglobin (Delta HbT) [coefficient = -3.825, standard error (s.e.) = 0.8353, z = -4.58, p < 0.001] and oxygenated hemoglobin (Delta HbO(2)) [coefficient = -4.6815, s.e = 0.6781, z = -6.90, p < 0.001] and a significant increase in deoxygenated hemoglobin (Delta HHb) [coefficient = 0.8192, s.e = 0.3034, z = 2.66, p < 0.01] across training. The results indicate that learning-related refinements in technical performance are mediated by temporal reductions in prefrontal activation.