9318061 Bloch Funds are requested to establish a multi-user confocal laser scanning microscope (CLSM) facility for the basic science departments at the University of Maryland at Baltimore. Such a shared instrument is not available. The applicants have been using fluorescence techniques to study cell structure and function, but progress in some aspects of their research has been limited by the resolution afforded by the optical microscope. With a confocal laser scanning microscope, the resolution in the x-y plane can be increased ~2-fold and in the z-axis, ~1.4 fold. The increased resolution provided by the confocal microscope should facilitate more definitive studies by the applicants of some important questions in modern cell biology. In particular, the confocal microscope will be used to study the organization of cellular membranes and how they change with differentiation or during endocytosis; how cell populations change during development of the brain; and how viruses and microbes enter cells and alter cell structure. All the applicants hold federal grants for their research, and several are recipients of grants from the NSF. The facility will include a CLSM system for data collection, and a separate workstation for image analysis. The CLSM system will consist of a Bio-Rad MRC-600 confocal laser scanning unit and an Olympus fluorescence microscope. The microscope will be configured to permit conventional epifluorescence and phase contrast for correlation with confocal images. The workstation, consisting of an IBM 486 running the same CoMOS program that controls the CLSM, is requested to maximize the availability of the CLSM for data collection. Both the CLSM system and the workstation will be provided with identical optical disk drives, to facilitate the transfer of image data acquired by the individual users. The hardware selected for this facility represents a reasonable balance between costs, our requirements for the highest res olution available for confocal microscopy, and the flexibility that is important for a multi-user facility. This equipment will be housed in the Department of Physiology, School of Medicine, University of Maryland at Baltimore. It will be administered by an interdepartmental Internal Advisory Committee consisting of the P.I. and two regular users, and supervised by Dr. Paul Luther, a co-investigator with experience in quantitative fluorescence techniques. The facility will be made available to all members of the university community, including students and postdoctoral fellows. Instruction in confocal microscopy will be provided by regular, practical training sessions and through an interdepartmental graduate course on imaging methodology. Realizing the importance of confocal microscopy to the research and teaching activities in the basic sciences, the Graduate School, University of Maryland, Baltimore, and the Dean of the School of Medicine have both committed matching funds to help establish the CLSM facility, should this grant be funded.
