Segmentation of three-dimensional images using non-rigid registration: Methods and validation with application to confocal microscopy images of bee brains

This paper describes the application and validation of automatic segmentation of three-dimensional images by non-rigid registration to atlas images. The registration-based segmentation technique is applied to confocal microscopy images acquired from the brains of 20 bees. Each microscopy image is registered to an already segmented reference atlas image using an intensity-based non-rigid image registration algorithm. This paper evaluates and compares four different approaches: registration to an individual atlas image (IND), registration to an average shape atlas image (AVG), registration to the most similar image from a database of individual atlas images (SIM), and registration to all images from a database of individual atlas images with subsequent fuzzy segmentation (FUZ). For each strategy, the segmentation performance of the algorithm was quantified using both a global segmentation correctness measure and the similarity index. Manual segmentation of all microscopy images served as a gold standard. The best segmentation result (median correctness 91 percent of all voxels) was achieved using the FUZ paradigm. Robustness was also the best for this strategy (minimum correctness over all individuals 84 percent). The mean similarity index value of segmentations produced by the FUZ paradigm is 0.86 (IND, 0.81; AVG, 0.84; SIM, 0.82). The superiority of the FUZ paradigm is statistically significant (two-sided paired t-test, P < 0.001).