Abstract--Due to the significant complexity of membrane morphology and the generally poor image quality in electron tomographic volumes, current automatic methods for segmentation of membranes perform poorly. Users must resort to manual tracing of recognized patterns on 2-D slices of the volume, a method that suffers from subjectivity and is very labor intensive, preventing quantitative analyses of tomographic data that require comparative analyses of many volumes. To overcome these limitations, we develop an automatic 3-D segmentation method that fully exploits the prior knowledge about the shape of the membranes as well as the 3-D information provided by the tomograms, and systematically combines this knowledge with the image data to improve segmentation results. The method is based on the watersnake framework. By mathematically reformulating the traditional watershed segmentation as an energy minimization problem, the watersnake inherits the many strengths of the watershed method wh...