Purpose: The location of cortical areas and visual fatigue related to stereoscopic vision were explored by two types of fMRI designs, namely block stimulation and event-related stimulation. The stimulations consist of 2D/3D images and different depths of the stereoscopic 3D images. Method: 20 normal subjects were randomly divided into the block group and the event-related group. Blood oxygenation level-dependent functional magnetic resonance imaging (Bold-fMRI) was performed in two groups. Functional data was preprocessed and statistically analyzed by SPM8. The result was reported by REST. Result: In the block stimulation group, compared to 2D image stimulation, 3D image stimulation results in more activated brain areas, including frontal lobes, occipital lobes and limbic lobes, especially in the frontal eye field (Brodmann Area 8, BA8) and middle occipital gyrus (BA18/19). In the event-related group, compared to 2D images, viewing 3D images causes significant activations in temporal lobe, mainly represented in BA19/13/31/37. Additionally, 3D image stimulation with the focus set at front depth can lead to the activation of more brain areas compared to the back depth, including inferior parietal lobule and posterior central gyms. Conclusion: The formation of the stereoscopic vision requires the collaboration of more brain areas, so that viewing stereoscopic videos for a long period may result in visual fatigue; meanwhile, the front depth of field can contribute to more activated brain areas than the back depth of field. As a parameter of stereoscopic images, it is valid to state that the depth of field may affect visual fatigue. (C) 2017 Elsevier B.V. All rights reserved.