Laser Scanning Confocal Microscope

This is a proposal for a multiparameter laser scanning confocal microscope system. The microscope system consists of a detachable confocal laser scanning unit with multi-line argon laser, dichroic beam splitter, and dual photon-counting detectors; upright and inverted microscopes with interchangeable optics; intensified CCD video camera for selecting and focusing fields with minimal photobleaching and photodamage; and on-stage environmental controllers. The microscope system will be used to examine tissues of living animals, single living cells, fixed cells, and tissue sections which have been labeled with multiple fluorescent probes. Primary uses will be determination of the 3- dimensional distribution of specific antigens in cells, visualization of complex neural networks, determination of lipid order and topography of cell membranes, and quantitation in 3- dimensions of ions (e.g., Ca and H+) and organelles (e.g., mitochondria, lysosomes, nuclear chronmatin) within living cells. The equipment will be utilized by 17 investigators on independent projects. Housed in the productive environment of the Video Microscopy Facility of the Laboratories for Cell Biology, the confocal microscope system will complement existing state-of-the- art instrumentation and will substantially extend our ongoing efforts applying digitized optical microscopy to problems of biology.