We developed a new minimal path segmentation method for mouse kidney MR images. We used dynamic programming and a minimal path segmentation approach to detect the optimal path within a weighted graph between two end points. The energy function combines distance and gradient information to guide the marching curve and thus to evaluate the best path and to span a broken edge. An algorithm was developed to automatically place initial end points. Dynamic programming was used to automatically optimize and update end points during the searching procedure. Principle component analysis (PCA) was used to generate a deformable model, which serves as the prior knowledge for the selection of initial end points and for the evaluation of the best path. The method has been tested for kidney MR images acquired from 44 mice. To quantitatively assess the automatic segmentation method, we compared the results with manual segmentation. The mean and standard deviation of the overlap ratios are 95.19%±0.03%. The distance error between the automatic and manual segmentation is 0.82±0.41 pixel. The automatic minimal path segmentation method is fast, accurate, and robust and it can be applied not only for kidney images but also for other organs.