This Immunology and Pathology Laboratory Core (Core B) will serve as a centralized facility for procedures common to all of the projects in the Program Project. This core will have the capacity to perform high throughput analysis of samples generated from animals and tissue culture. The core will perform multiplexing analysis of cytokine proteins from various samples generated from serum and cell culture. It will also perform RT-PCR for detection mRNA cytokines isolated from various tissues as needed. Finally, it will perform routine and specialized immunohistochemistry on tissue samples for examination of pathological specimens. It wil also assist with sample preparation from various tissues. All three projects will use this core for some element of their analysis. This Core will be the most heavily used for the PPG. Specifically this Immunology and Pathology core will: 1) Provide standardized methods for evaluating the level of cytokine proteins and/or their RNA in fluids and tissues collected from mice, human cells or primates. Methods will include multiplexing strategies for quantitatively detecting multiple cytokines in one sample, real-time PCR techniques for quantitative detection of cytokine RNAs, ELISpot methods to determine the number of cells collected from either organs or blood producing specific cytokines 2) Provide capability in increased labor capacity to prepare samples from mice and primates for histological and cellular analysis 3) Provide capability and capacity in assisting in disruption of lung cells and evaluation of cell phenotypes of cells recovered directly from organs or tissue culture by flow cytometry techniques. Other capabilities will be to perform intracellular cytokine staining for phenotyping cells recovered animal tissues and blood or potentially tissue culture as needed.
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