This is an application requesting purchase of a laser scanning confocal microscope for use in cell biological and neurobiological research projects at the UMDNJ-Robert Wood Johnson Medical School. Laser scanning confocal microscopy has become an increasingly valuable technique for high-resolution localization and reconstruction of fluorescently-tagged cellular proteins using immunofluorescence methods. Other fluorescent probes are also available for the localization and measurement of cellular calcium, pH and membrane potential. These advances are particularly important to the fields of cell biology and neurobiology because they allow increased understanding of the relationship of cellular constituents to cellular development and function. The equipment requested includes a Bio-Rad MRC-600 laser scanning confocal head and Zeiss Axioplan microscope. The equipment is to be used for the high-resolution localization and three-dimensional reconstruction of the distribution of cellular proteins by a group of highly-qualified and well-funded investigators. Seven major NIH-funded users are identified whose projects concern 1) cell surface proteins on neurons and glia; 2) the molecular basis of lysosomal enzyme targeting; 4) the genetic analysis of the role of tubulin and other proteins in mitosis; 4) role of cell proliferation in nervous system development; 5) the localization of mannose-6-phosphate receptors during neural development; 6) the modulation of cell structure by the adenovirus E1A product; 7) molecular analysis of myosin synthesis and assembly in muscle development. Each of these projects will be advanced significantly by the requested equipment. The research community on the Robert Wood Johnson Medical School/Rutgers University campus consists of over 100 NIH-funded investigators. There is no laser scanning confocal microscope available on this campus. Therefore, in addition to the Major Users, we have projected that approximately 30% of time will be available to these investigators to extend their research through the use of laser scanning confocal microscopy, three dimensional reconstruction and mathematical analysis of the co-localization of cellular proteins. We have included abbreviated descriptions from five of these to indicate the value of the instrument to the research community.
