The objective of the Quantitative Molecular Pathology Core (D), to be located in the Departments of Anatomy and Psychiatry at Charing Cross and Westminster Medical School, will be to provide rapid, reproducible, and reliable analysis of the expression of molecules which are of interest to the principle investigators of this Program Project. In particular, this Core will determine (i) the levels of specific proteins and messenger RNAs (mRNAs); and (ii) the number and spatial orientation of cells or components expressing these molecules. Tissue sections or x-ray films (derived from studies of human or experimental animal brain specimens performed by Project Investigators) will be analyzed such that information regarding the number of relevant cells or structures per brain area as well as the levels of the specific proteins and mRNAs relative to total proteins and ribosomal or polyadenylated mRNAs in specific brain regions will be available to Project Investigators requesting services from this Core. The determinations required to provide these services will be performed using specific image analysis protocols designed by the Director of this Core. The image analysis system presently in place and the relatively inexpensive freestanding workstation requested in this application will provide a dedicated image analysis system (Seescan PLC, Cambridge, UK) of sufficient capacity and quality to provide appropriate analysis of the large volume of material that will he generated in the Projects of this Program. The analysis protocols to be used in this Core have been designed to supersede more traditional visual inspection strategies for examination and evaluation of neuropathological materials. In order to determine the extent to which a pathological process has occurred, more traditional methods involve complex and skilled procedures performed by an individual making subjective interpretive decisions as microscopic images are visualized. Using the system and algorithms devised and proposed here, this Core will provide standardized analysis of material derived from Project investigation by eliminating subjective elements inherent to manual techniques, especially when these techniques are performed by several individuals. In addition, the availability of this Core for analysis of the large volume of material to be generated by the Projects of this Program will reduce a laborious, time consuming and thus expensive endeavor to a manageable and economically feasible level.
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