In clinical research using medical images, the image segmentation is one of the most important processes. Especially, the hippocampal atrophy is helpful for the clinical Alzheimer diagnosis as a specific marker of the progress of Alzheimer. In order to measure hippocampus volume exactly, segmentation of the hippocampus is essential. However, the hippocampus has some features like relatively low contrast, low signal-to-noise ratio, discreted boundary in MRI images, and these features make it difficult to segment hippocampus. To solve this problem, firstly, We selected region of interest from an experiment image, subtracted a original image from the negative image of the original image, enhanced contrast, and applied anisotropic diffusion filtering and gaussian filtering as preprocessing. Finally, We performed an image segmentation using two level set methods. Through a variety of approaches for the validation of proposed hippocampus segmentation method, We confirmed that our proposed method improved the rate and accuracy of the segmentation. Consequently, the proposed method is suitable for segmentation of the area which has similar features with the hippocampus. We believe that our method has great potential if successfully combined with other research findings.