This proposal requests funds to assist in the purchase of a Confocal Laser Scanning Microscope System (CLSM). The enormous and rapidly expanding battery of antibodies and of other fluorescent probes has opened the way to a greatly expanded use of fluorescent microscopy. Unfortunately, the thickness and light scattering occurring in many biological specimens seriously compromises the sensitivity, clarity and resolution of even the best epi-fluorescent images. CLSM overcomes these limitations and produces images of great clarity. This instrumentation will serve the needs of biological scientists at the University of Wyoming (UW) whose work focuses on cell architecture and cellular dynamics, organelle structures and their spatial and temporal relationships and the localizations/functions of cell and tissue antigens. The proposal is submitted by a major user group representing four UW departments: Botany, Molecular Biology, Psychology, and Zoology and Physiology. The system will be available to all UW researchers who have legitimate needs and will operate as a bona fide University instrumentation center. The acquisition of this state-of-the-art instrumentation and the establishment of a CLSM center will offer investigators the opportunity to remain competitive in cell/molecular biology and neuroscience, foster research collaborations and provide training for both undergraduate and graduate students. The CLSM system will initially be used in support of the following projects: Chromosomal Studies in Bromeliaceae (G. Brown, Botany), The Role of Bombesin-like Peptides in Neural Physiology (F. Flynn, Psychology), Applications of Confocal Laser Microscopy to Research on Nuclear Phenomena in Ciliate Protozoa (R. Jenkins, Zoology), Probing Plant-Fungal Interactions: The Form and Physiology of Ectomycorrhizal Fungal Linkages with Plant Roots (S. Miller, Botany), Effects of Heat on Development: Use of the Confocal Microscope to Follow Assembly of Actin Fibers and Localizati on of Integrins Following Heat Shock (N. Petersen, Molecular Biology), Applications of Confocal Microscopy to Research in Behavioral Neuroendocrinology (J. Rose, Psychology), Confocal Microscopy in Studies of DNA Escape from Mitochondria to the Nucleus (P. Thorsness, Molecular Biology), Laser Confocal Microscopy in Studies of Leaf Structure Function: Do Epidermal Lens Cells Play a Role in Photosynthesis? (T. Vogelmann, Botany). In addition, the CLSM system will be used as a teaching tool within laboratory based courses within the four departments as well as for closely supervised graduate and undergraduate research. The total project cost is $253,710 of which the University of Wyoming has pledged a 49% match. NSF FORM 1358 (1/94) 3
