Geometric approach to segmentation and protein localization in cell cultured assays

Abstract

Cell-based fluorescence imaging assays are heterogeneous requiring collection of a large number of images for detailed quantitative analysis. Complexities arise as a result of variation in spatial nonuniformity, shape, overlapping compartments, and scale. A new technique and methodology has been developed and tested for delineating subcellular morphology and partitioning overlapping compartments at multiple scales. This system is packaged as an integrated software platform for quantifying images that are obtained through fluorescence microscopy. Proposed methods are model-based, leveraging geometric shape properties of subcellular compartments and corresponding protein localization. From the morphological perspective, convexity constraint is imposed to delineate, partition, and group nuclear compartments. From the protein localization perspective, radial symmetry is imposed to localize punctate protein events at sub-micron resolution. The technique has been tested against 196 images that were generated to study centrosome abnormalities. Computed representations are evaluated against the ground truth annotation for comparative analysis. © Springer-Verlag Berlin Heidelberg 2005.