Segmentation method for three-dimensional visualization of microscopic objects imaged with a confocal laser scanning microscope

Segmentation, or separating an image into distinct objects, is the key to creating 3-D renderings from serial slice images. This is typically a manual process requiring trained persons to tediously outline and isolate the objects in each image. We describe a template-based semiautomatic segmentation method to aid in the segmentation process and 3-D reconstruction of microscopic objects recorded with a confocal laser scanning microscope (CLSM). The simple and robust algorithm is based on the creation of a userdefined object template, followed by automatic segmentation of the object in each of the remaining image slices. The user guides the process by selecting the initial image slice for the object template, and labeling the object of interest. The algorithm is applied to mathematically defined shapes to verity the performance of the software. The algorithm is then applied to biological samples, including neurons in the common housefly. It is the quest to further understand the visual system of the housefly that provides the opportunity to develop this segmentation algorithm. Further application of this algorithm may extend to other registered and aligned serial section datasets with high contrast objects. (c) 2006 SPIE and IS&T.