Advanced Imaging & NanoString Core Research on human skin diseases has been hampered by limitations on the number of studies that can be done on formalin fixed paraffin embedded (FFPE) specimens, the most frequent type generated in the course of clinical care, by clinical trials and available in specimen archives. The purpose of this Core is to accelerate human skin disease research by providing access to two transformative techniques, six color immunostaining, spectral imaging and automated cell analyses and NanoString-based analyses, which taken together allow comprehensive evaluation of protein and gene expression in FFPE specimens. Tyramide amplification based serial immunostaining allows six color labeling of FFPE specimens with no antibody species interference, long term signal stability and minimal quenching. We provide multi spectral imaging on the cutting edge Perkin- Elmer Mantra imaging system, providing image deconvolution and automated cell analyses. The techniques allow automatic identification, counting, and measurement of fluorescence signals in other channels for individual cell types in the section, yielding the equivalent of flow cytometry data from immunostained tissue sections. The Core also provides validated antigen retrieval and staining protocols, validated antibody panels, and staining and image acquisition services that make these techniques rapidly accessible to researchers at any institution. NanoString analysis provides quantitative measurement of up to 800 DNA or RNA targets in a single test run and is the first nucleic acid analysis technique that works well in FFPE tissues. The Core provides access to a cutting-edge n-Counter instrument, as well as sample preparation and data acquisition services that make these analyses accessible to outside investigators. This Core is run by renowned dermatopathologist Dr. Martin Mihm, who will provide consultative assistance to Center users to enhance their understanding of human skin disease. The research community potentially includes any individual wishing to carry out human skin disease research. We have included projects from 24 investigators who wish to utilize Center services, 23 of whom would like to utilize the Advanced Imaging & NanoString Core for their projects. 15 are from outside institutions, nine have never worked in human skin disease before and seven have worked primarily in mouse models previously, with little or no prior work in humans. Eight of these projects are described in detail in this component and other projects are described fully in the two other Resource Core portions of the application. In summary, the Advanced Imaging & NanoString Core provides access to two powerful new techniques that allow comprehensive analysis of protein and gene expression in FFPE samples, with the goal of accelerating human skin disease research.
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