Neuroticism is a core personality trait that profoundly affects how individuals interpret and interact with their environment. Understanding neuroticism at a neurobiological level will be an important step toward identifying novel vulnerability factors for psychiatric illnesses such as depression and anxiety. Along these lines, recent work has identified neural activation patterns within the right anterior insula that correlates with an individual's degree of neuroticism. The present study aims to further characterize the circumstances under which neuroticism modulates insular activity. Sixteen healthy participants underwent functional magnetic resonance imaging while playing a card game with varying degrees of outcome uncertainty. Activation within the bilateral anterior insula was found during all decisions, irrespective of uncertainty. However, a significant positive correlation between neuroticism and anterior insula activity was found only during 'certain decisions' (i.e. situations where the most probable outcome was clearly evident). Moreover, an increase in the right anterior insula activity during certain decisions was related to a behavioral mirroring effect such that the response latency for certain decisions approached the response latency for uncertain decisions. These findings suggest that increasing levels of neuroticism modulate neural activation in such a way that the brain interprets certainty as uncertain.