Automated local bright feature image analysis of nuclear proteindistribution identifies changes in tissue phenotype Page: 2 of 28
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The organization of nuclear proteins is linked to cell and tissue phenotypes. When cells arrest
proliferation, undergo apoptosis, or differentiate, the distribution of nuclear proteins changes.
Conversely, forced alteration of the distribution of nuclear proteins modifies cell phenotype.
Immunostaining and fluorescence microscopy have been critical for such findings. However,
there is an increasing need for quantitative analysis of nuclear protein distribution to decipher
epigenetic relationships between nuclear structure and cell phenotype, and to unravel the
mechanisms linking nuclear structure and function. We have developed imaging methods to
quantify the distribution of fluorescently-stained nuclear protein NuMA in different mammary
phenotypes obtained using three-dimensional cell culture. Automated image segmentation of
DAPI-stained nuclei was generated to isolate thousands of nuclei from three-dimensional
confocal images. Prominent features of fluorescently-stained NuMA were detected using a novel
local bright feature analysis technique, and their normalized spatial density calculated as a
function of the distance from the nuclear perimeter to its center. The results revealed marked
changes in the distribution of the density of NuMA bright features as non-neoplastic cells
underwent phenotypically normal acinar morphogenesis. In contrast, we did not detect any
reorganization of NuMA during the formation of tumor nodules by malignant cells. Importantly,
the analysis also discriminated proliferating non-neoplastic cells from proliferating malignant
cells, suggesting that these imaging methods are capable of identifying alterations linked not
only to the proliferation status but also to the malignant character of cells. We believe that this
quantitative analysis will have additional applications for classifying normal and pathological
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Knowles, David; Sudar, Damir; Bator, Carol & Bissell, Mina. Automated local bright feature image analysis of nuclear proteindistribution identifies changes in tissue phenotype, article, February 1, 2006; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc890050/m1/2/: accessed June 24, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.