The objective of establishing this centralized laboratory facility is to make the best use of the Center's resources and to avid redundance in equipment and personnel. In this core unit, we propose to establish two independent laboratories. They are a Protein Analysis Laboratory and Histology Laboratory. The major responsibility of this centralized core facility is to provide technical service and support to four proposed research projects. Since all procedures in these laboratories require precision and repetitive manipulations, they can be best carried out by centralized and experienced personnel. The Protein Analysis Laboratory will include analysis of complex protein mixtures from specimens by high- pressure liquid chromatography (HPLC) and two-dimensional gel electrophoresis (2-D). The HPLC will be used to separate complex mixtures of proteins into fractions of biologically active proteins for further analysis. 2-D electrophoresis will include the ISO-DALT System to separate proteins by isoelectric points in the first dimension and molecular weights in the second dimension. In addition to silver-staining to visualize the protein spots, the techniques of Western blotting and fluorography will be used. A computer-based image analysis system will also be used in qualitative and quantitative assessment of the resulting two-dimensional protein profiles. It is anticipated that approximately 400 gels will be processed by this laboratory each year (100 gels for each project). this capacity represents half of the operating capacity of the laboratory. The Histology Laboratory will be responsible for processing of all specimens generated by the four research projects for light microscopy. Besides the routine H&E sections, the laboratory will conduct immunohistochemistry, autoradiography, and quantitative analysis by a computer-based image analysis system.
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