Quantitative Representation Of Three-Dimensional Cell Culture Models

Three-dimensional mammary cell culture models offer new opportunities for the development of computational techniques for segmentation, localization, and multicellular organization. Under normal conditions, these assays form a symmetrical, hollow structure, which is necessary for their functional operation. Often, the nuclear compartments are labeled, which provides context for quantitative protein localization or colony structure through fluorescent microscopy. These colonies are first delineated from the background using the level set method. Within each colony, nuclear regions are then bounded by their center of mass through iterative radial voting, and a local neighborhood for each nucleus is established through Voronoi tessellation. Finally, the level set method is applied again within each Voronoi region to delineate the nuclear compartment. The paper concludes with the application of the proposed method to a set of experimental data demonstrating a stable solution when iterative radial voting and level set methods are used synergistically. Furthermore, segmented colonies are characterized for architectural changes as as a result of ionizing radiation.