© 2021 Emre Mülazimoǧlu et al.To mitigate dictionary attacks or similar undesirable automated attacks to information systems, developers mostly prefer using CAPTCHA challenges as Human Interactive Proofs (HIPs) to distinguish between human users and scripts. Appropriate use of CAPTCHA requires a setup that balances between robustness and usability during the design of a challenge. The previous research reveals that most usability studies have used accuracy and response time as measurement criteria for quantitative analysis. The present study aims at applying optical neuroimaging techniques for the analysis of CAPTCHA design. The functional Near-Infrared Spectroscopy technique was used to explore the hemodynamic responses in the prefrontal cortex elicited by CAPTCHA stimulus of varying types. The findings suggest that regions in the left and right dorsolateral and right dorsomedial prefrontal cortex respond to the degrees of line occlusion, rotation, and wave distortions present in a CAPTCHA. The systematic addition of the visual effects introduced nonlinear effects on the behavioral and prefrontal oxygenation measures, indicative of the emergence of Gestalt effects that might have influenced the perception of the overall CAPTCHA figure.