This proposal requests funds for a JEOL 1230 Transmission Electron Microscope (TEM) and a BOC Edwards Auto306 Vacuum Evaporator. These instruments will be located in the University of Iowa Central Microscopy Research Facility. The JEOL 1230 is being requested because the facility currently has just one TEM, a Hitachi H-7000 purchased with university funds in 1987. During the past 12 months, 78 faculty labs have used this instrument to support their research. 60 of these investigators have NIH funded research grants. 8 students each semester are enrolled in an Electron Microscopy Techniques course and use this instrument. Investigators have to sign up for the Hitachi TEM 2 to 3 weeks in advance. A number of operators are using the TEM during evenings and weekends. There is no guarantee that the instrument will be functional at a scheduled period because of unexpected downtime for maintenance. This results from a combination of very heavy use, the age of the Hitachi, which is a design introduced in 1981, and the low level of expertise of service personnel. Investigators also need low dose imaging, automated stage controls, tomography, digital imaging and Cryo-TEM to study their samples. The Hitachi TEM lacks all of these modern capabilities. The BOC Edwards Vacuum Evaporator will replace a 30 year old Varian unit in the facility. Repair parts have been unavailable for over 15 years; it suffers from poor vacuum and inconsistent evaporation of carbon and metals. It is critical that carbon coated grids for thin-sections and carbon thin films for negative staining, metal coating and Cryo-TEM be very-thin and low contrast. The JEOL TEM and the BOC Edwards evaporator will be conveniently located in room 84 and 92 respectively in the Eckstein Medical research Building as part of the University of Iowa Central Microscopy Research Facility. Administration and maintenance, as well as high-level technical assistance and training, will be provided by experienced personnel within the facility. Institutional support is strong with $61,234.00 being provided towards purchase of the instruments, $55,133.00 in full salary support for a research assistant and $10,000.00 to partially fund the annual service contract for the JEOL TEM on a continuing basis. Welsh will study airway epithelial cell biology, airway infections and gene transfer. Apicella plans to investigate the pathogenesis of bacteria. Dailey will examine synapse development and injury in brain. McCray will be involved with gene transfer using lentiviral vectors. Tobacman will study the structure of short actin filaments. Zabner will examine the morphology of Adeno-associated virus.
