Flux-based 3D segmentation of keratin intermediate filaments in confocal laser scanning microscopy

We propose an optimally oriented flux-based method for the 3D segmentation of keratin intermediate filaments (KFs) in images acquired using fluorescence confocal laser scanning microscopy (CLSM). KFs are elastic cables forming a complex scaffolding within epithelial cells that is involved in many basic cell functions. Segmentation of KFs to generate a graph representation of the KF network therefore is a key component for analyzing the KFs´ dynamic and biomechanical properties. Our segmentation method follows the principle of line enhancement, thresholding and centerline extraction to yield a one-voxel-wide centerline as result. The method is quantitatively evaluated using synthetic data generated by simulating the imaging process in CLSM. It is compared to segmentation results achieved with well-known vesselness measures. The results show that despite limited image quality in CLSM a high degree of precision is achieved.