This proposal seeks to acquire a point-scanning confocal fluorescence microscope for the Department of internal medicine at Yale University School of Medicine. The ongoing renaissance in the use of light microscopy for cell and molecular biology and physiology has been driven in large part by the development of specific fluorescent probes of cell structure and function, coupled with confocal fluorescence microscope systems that extend our abilities to visualize and quantitate such probes in cells and tissues. Laser scanning confocal microscopy, because of its ability to optically section cells and tissues, provides a tool with the spatial resolution needed to define structure in live and fixed cells alike, along with the temporal resolution to follow dynamic processes in living cells. Specific projects that would take immediate advantage of this equipment include studies of invasion and intracellular trafficking of the spirochete that causes Lyme disease, Borrelia burgdorferi, examination of the inflammation caused by Ehrichiosis and other tick-borne diseases, the molecular basis for pancreatic secretion and pancreatitis, alterations in subcellular calcium signaling that occur as a result of genetic defects in polycystic kidney disease, and endocytosis of Toxoplasma and other protozoans. Moreover, this confocal microscope would be the only such instrument in a new research building that will house 57 NIH-supported investigators who are members of the Department of Medicine. Thus this confocal microscope would be the basis of a core facility to assure that these investigators will have adequate access to state-of-the-art imaging techniques.
