(from application): The goal of this proposal is to stimulate and facilitate cutaneous research by providing cost-effective access to, and expert help in, the analysis of samples of normal and diseased skin. This will be achieved through a Tissue, Cellular and Molecular Analysis Core (TCMAC) which subsidizes the preparation and analysis of tissue sections, proteins, and nucleic acids derived from experimental tissues provided by investigators, or from banked samples of tissue, serum, and nucleic acids maintained by or accessible to the TMAC facility. Sample sources in hand consist of over 150,000 archival biopsies of diseased skin and serum samples that were submitted to the Yale Dermatopathology Service, and that are cross-referenced by diagnosis in a relational database. Serum samples, tissue sections, and nucleic acids extracted from multiple cases of common and rare diseases can be provided to researchers from this tissue bank without patient identification or specific Human Investigation Committee approval, because they represent the residua of material submitted for diagnosis.
The specific aims are to provide a range of services including, but not limited to: 1) subsidized preparation of unstained tissue sections and sections stained using a complete range of techniques including standard and extraordinary histochemical stains, immunohistochemical stains, and immunofluorescence stains; 2) subsidized two color flow cytometry including help with preparation of samples and analysis of results; 3) genomic DNA, total RNA, and single-stranded random primed cDNA libraries that are derived from fresh and archival samples of normal and diseased skin, and that are suitable for polymerase chain reaction (PCR) analysis; 4) antibody and nucleic acid probes and primers relevant to skin disease research; 5) assistance with the preparation and use of nucleic acid probes including isotopically and non-isotopically labeled oligonucleotide, cDNA, and cRNA probes, and assistance with southern and northern blotting and in situ hybridization; 6) advice and assistance in the design, construction and analysis of transgenes including the provision of tested skin specific promoter constructs and the detection of transgene integration and transcription; and 7) expertise in the use of all of the above described techniques and materials for the analysis of gene expression at all levels, from mRNA to protein, in whole tissue and individual cells, including morphologic analysis. Funding of this Core will encourage investigation of skin diseases by providing appropriate samples to researchers, including those whose primary area of interest is not the skin, and by providing economics of scale and the highest levels of expertise to investigators of all backgrounds.
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