Aim: This study aimed to evaluate the physiological correlates of cognitive load in laparoscopic surgery by assessing heart rate (HR), heart rate variability (HRV), pupil dilation, galvanic skin response (GSR), and electroencephalography (EEG) activity among surgeons with varying experience levels. Materials and Methods: A prospective observational study was conducted on 100 laparoscopic surgeons, categorized into novice (n=35), intermediate (n=40), and expert (n=25) groups based on their surgical experience. Participants performed standardized laparoscopic tasks in a controlled environment while physiological parameters were continuously recorded. HR and HRV were measured using ECG devices, pupil dilation with an eye tracker, GSR with skin conductance sensors, and EEG with a portable system. The NASA Task Load Index (NASA-TLX) was used to assess perceived workload, while surgical performance was evaluated using the Global Operative Assessment of Laparoscopic Skills (GOALS). Results: Novices exhibited significantly higher HR (110.4 ± 10.1 bpm vs. 92.1 ± 7.5 bpm, p < 0.001), lower HRV (RMSSD: 32.1 ± 8.2 ms vs. 45.7 ± 6.9 ms, p < 0.05), increased pupil dilation (6.2 ± 0.8 mm vs. 4.9 ± 0.5 mm, p < 0.001), and elevated GSR (2.5 ± 0.5 μS vs. 1.2 ± 0.3 μS, p < 0.001) compared to experts. EEG analysis revealed higher frontal theta power (4.5 ± 1.2 µV² vs. 3.2 ± 0.9 µV², p < 0.01) and lower alpha power (2.1 ± 0.9 µV² vs. 3.1 ± 0.7 µV², p < 0.05) in novices, indicating increased cognitive effort. Subjective workload (NASA-TLX) was significantly higher in novices (72.3 ± 8.5 vs. 47.2 ± 6.4, p < 0.001), while experts achieved better GOALS scores and shorter task completion times. Strong correlations were observed between NASA-TLX and physiological markers such as HR increase (r = 0.72, p < 0.001), pupil dilation (r = 0.65, p < 0.001), and GSR (r = 0.59, p < 0.01). Conclusion: Novice surgeons exhibit higher cognitive load, greater physiological stress responses, and lower surgical efficiency than experienced surgeons. These findings highlight the potential of real-time physiological monitoring to assess cognitive load during surgical training. Integrating stress management and cognitive training techniques could enhance skill acquisition and optimize performance in laparoscopic surgery.