Sustained attention is the ability to keep focused and vigilance for long time in external stimulation, which was crucial in safe-critical human-machine system. While the ability of sustained attention will decline because of mental fatigue, even lead to serious accidents in fatigue state. Therefore, it is of great significance to explore the impact of fatigue on sustained attention. Functional Near-Infrared Spectroscopy (fNIRS) can measure cerebral hemoglobin in order to reflect cognitive function indirectly. In previous related fatigue studies, monotonous and long-time CPT (continuous performance test task) was often used to explore the performance change and brain activity, but the effect of time on task (TOT) was always involved. In this study, in order to avoid the TOT effect, the sustained attention task and fatigue task were separated. It was adopted in the study that the modified continuous performance test (CPT) was chosen as the sustained attention task and verbal 2-back task as the fatigue induced task. The fNIRS signals were extracted from 10 channels in the prefrontal cortex (PFC) from 20 healthy subjects. Studies found that cerebral lateralization increased significantly from alert to fatigue state in sustained attention task. Besides, Average oxyhemoglobin (HBO) of PFC increased significantly from alert to fatigue task, and the spatial pattern of activity of oxyhemoglobin also changed, which c be sensitive features to fatigue detection. © 2017 SPIE.