The purpose of Primary Cell, Tissue Processing and Imaging Core (Core C) is to provide centralized and expert handling and processing of primary cells, tissues, and to serve as an imaging facility. This Core will be in charge of establishing and maintaining techniques for culturing primary human cells, including airway epithelial cells, and will provide molecular cellular, histological and ultrastructural information for each of the experimental projects. It is highly beneficial to the Center that these activities be performed in a centralized fashion, as it would be difficult and inefficient to run these highly specialized functions by separate laboratories. The Specific Aims of Core C are: Aim 1: The Core will provide expert, centralized initial processing and characterization of human samples. It will transform these samples into primary cells in culture. The Core will develop a cryogenic bank of primary cells, ensure appropriate storage, inventory, and coordinate their distribution to Center investigators. Aim 2: The Core will provide molecular, cellular, histological and ultrastructural information for each of the experimental projects. Core C will perform primary cell culturing, histology, immunohistochemistry, fluorescence microscopy deconvolusion microscopy, live cell imaging, confocal microscopy and electron microscopy. Additional services, including three-dimensional image analysis of fluorescently labeled cells using deconvolution microscopy, will also be available. The facilities available for the Core are all located within close proximity of the investigators'laboratories on the 5th and 6th floor of Baylor's main building. The Core has fluorescence and light microscopy with computer-assisted image analysis and deconvolution microscopy with three-dimensional image analysis capability. In addition, physically located within the Center is the Seymour Lieberman Electron Microscopy Laboratory, which is a dedicated facility for transmission electron microscopy and morphometric analysis. Specialized techniques will be used to localize specific proteins in primary cells, cell lines, and lung tissues. For monolayer cell culture analysis, light microscopic level using immunohistochemistry and, in cases where fluorescent antibody detection is used or when fluorescent fusion proteins are utilized, deconvolution microscopy will be employed in conjunction with computerassisted three-dimensional image reconstruction and analysis. Confocal microscopy will be used for thick tissue sections, for some live cell imaging experiments and for some of the experiments requiring laser, e.g., fluorescence recovery following photobleaching. In addition, Core personnel will carry out initial tissue processing and analysis.
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