Driver fatigue is a common cause of car accidents. Thus, the objective detection of driver fatigue is a first step toward the effective management of fatigue-related traffic accidents. Here, we investigated the effects of driving time, a common inducer of driver fatigue, on the dynamics of fixational eye movements. Participants drove for 2 h in a virtual driving environment while we recorded their eye movements. Microsaccade velocities decreased with driving time, suggesting a potential effect of fatigue on microsaccades during driving.