Fundamentals of Laparoscopic Surgery (FLS) is a prerequisite for board certification in general surgery in the USA. In FLS, the suturing task with intracorporeal knot tying is considered the most complex. Transcranial direct current stimulation (tDCS) of the dorsolateral prefrontal cortex (PFC) has been shown to facilitate FLS surgical skill acquisition where 2mA tDCS for 15min with the anode over F3 (10/10 EEG montage) and cathode over F4 has improved performance score in an open knot-tying task. Since PFC has a functional organization related to the hierarchy of cognitive control, we performed functional near-infrared spectroscopy (fNIRS) to investigate PFC activation during the more complex FLS suturing task with intracorporeal knot tying. We performed fNIRS-based analysis using AtlasViewer software on two expert surgeons and four novice medical students. We found an average cortical activation mainly at the left medial PFC across the experts, while the average cortical activation across the novices was primarily at the left pars opercularis of the inferior frontal gyrus and ventral premotor cortex, inferior parietal lobule, and supramarginal gyrus. Here, the average cortical activation across the novices included not only the cognitive control related brain regions but also motor control complexity related brain regions. Therefore, we investigated the feasibility to stimulate motor complexity related PFC regions using ROAST software and identified a 4x1 high-definition (HD) tDCS montage.Clinical Relevance—This shows the feasibility of fNIRS-guided tES to individualize electrode montage that may facilitate FLS surgical training in our future studies.