Jun Wang, Xiaobo Zhou, Pamela L. Bradley, Shih-Fu Chang, Norbert Perrimon, Stephen T.C. Wong. Cellular Phenotype Recognition for High-Content RNAi Genome-Wide Screening. Journal of Biomolecular Screening, 13(1):29-39, February 2008.
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Genome-wide, cell-based screens using high-content screening (HCS) techniques and automated fluorescence microscopy generate thousands of high-content images that contain an enormous wealth of cell biological information. Such screens are key to the analysis of basic cell biological principles, such as control of cell cycle and cell morphology. However, these screens will ultimately only shed light on human disease mechanisms and potential cures if the analysis can keep up with the generation of data. A fundamental step toward automated analysis of high-content screening is to construct a robust platform for automatic cellular phenotype identification. Here, we present a framework, consisting of microscopic image segmentation and analysis components, for automatic recognition of cellular phenotypes in the context of the Rho family of small GTPases. To implicate genes involved in Rac signaling, RNA interference (RNAi) was used to perturb gene functions, and the corresponding cellular phenotypes were analyzed for changes. The data used in our experiments are high-content, 3 channels, fluorescence microscopy images of Drosophila Kc167 cultured cells stained with markers that allow visualization of DNA, polymerized actin filaments and the constitutively activated Rho protein RacV12. The performance of this approach was tested using a cellular database that contained more than 1,000 samples of three predefined cellular phenotypes, and the generalization error was estimated using a cross validation technique. Moreover, we applied this approach to analysis the whole high-content fluorescence images of Drosophila cells for our further HCS based gene function analysis
@article{wj:jbs2008,
Author = {Wang, Jun and Zhou, Xiaobo and L. Bradley, Pamela and Chang, Shih-Fu and Perrimon, Norbert and T.C. Wong, Stephen},
Title = {{Cellular Phenotype Recognition for High-Content RNAi Genome-Wide Screening}},
Journal = {Journal of Biomolecular Screening},
Volume = {13},
Number = {1},
Pages = {29--39},
Month = {February},
Year = {2008}
}
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