This proposal is for funding for a BioRad DVC-250 laser-scanning confocal microscope, together with associated camera, computer and printer equipment. This equipment will be used for imaging both living and fixed biological specimens, at high resolution, and in three dimensions. This rapidly maturing technology is now in very wide use, and is particularly suited to visualizing multiple antigens or stains, simultaneously, and their digital analysis. With the equipment system requested, we will be able to acquire microscopic images, process them with a computer, and print them out, at publication quality. While there are a number of other microscopes available on this campus (compound microscopes, Electron microscopes, etc.), there is no confocal microscope. The USC medical school houses a confocal system, but this is not available to labs on this campus. Two of the proposed major users now use confocal systems at other sites. Ko uses a system at the City of Hope Hospital (20 miles distant), which is not available to others. Warrior uses a system at U.C. Irvine (50 miles distant), which is available, but charges a $100/hour user fee. In short, the biological researchers here do not have regular, or open access to this important technology. We have examined three different confocal microscopes: the Nikon K2Bio, BioRad DVC-250, and the Meridian, and we have determined that the BioRad offers the best compromise of performance and cost, for our purposes. This institution (USC), has undertaken to provide 30% of the total cost of this equipment, as a cost share. This equipment system will be used by a group of Biological research labs at the University Park campus, of the University of Southern California (in Los Angeles). This group consists of five major users (Moses, Warrior, Tower, Herrera and Ko, the principal investigators listed), and five minor users (Bottjer, Manahan, McFall-Ngai, Thompson and Watts). All of the major users are currently supported by federal gr ants. While our research interests are somewhat disparate we all share a strong interest in using the confocal microscope to visualize structures and antigens in three dimensions in biological specimens. The Moses lab studies pattern formation and cell fate determination in the Drosophila compound eye. They will use the confocal system to colocalize gene products (such as Drosophila homologs of Transforming Growth Factor a, and its putative receptor), and to examine double and triple stained genetic mosaic tissues. The Warrior lab studies inter and intra-cellular movements in the developing Drosophila embryo. They will use the equipment to study germ cell migration, and the functions of the schnurri and DnudC genes in early development. The Tower lab studies development of the Drosophila chorion, and the regulation of DNA replication. They will use the system to quantitate DNA levels in the developing follicle cells. The Herrera lab studies developmental plasticity in frog neuromuscular junctions, and the action of steroid hormones on this structure. They will use the system to study the junction structures in living specimens, in which their function, and response to hormones, can be tested. The Ko lab studies neuro transmitter release and the function of the presynaptic terminal. They will use this equipment to explore the behavior and morphology of voltage coupled Calcium channels, using specifically binding toxins.
