The purpose of this Shared Instrumentation Grant application is to establish an image analysis facility which will function as a shared resource under the auspices of the Duke University Comprehensive Cancer Center and the Duke Center for Aging. The primary task of this facility will be to perform quantitative analyses of 2-dimensional (2-D) gel electrophoresis images. 2-D gels are being widely used at this Medical Center in a broad range of projects, including efforts to understand AIDS, growth regulation, cystic fibrosis, mechanisms of aging and hormone action. Expertise for running extraordinarily high resolution (""""""""giant"""""""") 2-D gels exists at Duke University. These 2-D gels can resolve up to 5,000 proteins from complex biological samples in a manner suitable for measuring relative abundance, synthetic rate or post-translational modification. Proteins of interest can also be purified and partially sequenced from these gels. This information will allow the isolation of the cDNAs or genes coding for these proteins, using techniques and equipment available in other Cancer Center Shared Resources. Because of the resolving power of the giant gels, experiments based on this technique can yield insights unobtainable by other means. However, the complexity of the resulting data means that much information is lost unless the gel images can be analyzed, quantitated and the output stored in a powerful database format which allows investigators to track changes in potentially thousands of gene products across a series of experiments. In addition to providing these essential 2-D gel analysis functions, the proposed image analysis facility will also be used for other applications entailing quantitation of less complex images, such as those generated by 1-dimensional gel electrophoresis and autoradiograms, Western (immuno-) blots, thin layer chromatography including such applications as CAT (chloramphenicol acetyl transferase) assays to monitor gene expression, restriction-fragment length polymorphism analysis, slot-, dot-, northern and Southern blots for the quantitation of nucleic acids, and even DNA sequencing gels. Experience with similar image analysis facilities at other medical centers indicates that these additional applications prove to be tremendously valuable to the general research community. The proposed facility will include fully developed software which will enable large numbers of investigators to perform state-of-the-art analyses of their data. Furthermore, the hardware platform it will also permit interested investigators to work on improving computational aspects of image analysis procedures as part of an ongoing collaboration with Cal Tech and the Jet Propulsion Laboratory. Local commitment of funds by the Cancer Center at Duke to support operating costs means that user fees will be modest. No comparable instrumentation is available in North Carolina.
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