Funds requested in this proposal will be used to establish a Laser Scanning Confocal Microscope facility (LSCM) within the Division of Central Research Laboratories at Rhode Island Hospital. The Nikon PCM 2000 System will significantly extend the research capabilities of a major user group comprising of six NIH-funded investigators by providing cutting-edge instrumentation and technologies which will be utilized to study living and fixed cells and tissues using a range of contrast modes and fluorescent probes. A diverse array of tissues and isolated/cultured cell types will be analyzed including liver, brain, spleen, intestinal mucosa, and cells of the reticuloendothelial system (among others). The Nikon PCM 2000 System incorporates: (a) the Nikon Eclipse E 800 Research Upright Microscope and the Research Nikon TE-200 Inverted Microscope (configured with bright field phase contract, differential interference contrast, and fluorescence microscopy optics), (b) multiple wave length excitation capability by means of a fiber optically coupled, Argon Ion/HeNe laser illumination source; (c) multiple wave length detection through a high resolution, two channel fluorescence detector system; (d) including hard disk archiving capability; (e) data transfer via Ethernet to adjacent and/or remote pre-existing image analysis workstations; and (f) three dimensional reconstruction software. Specific applications made possible by this instrumentation include: (a) detection and discrimination of fluorescent probes in single-, double-, or triple-labeled samples; (b) monitoring of intracellular activities with vital or pH sensitive fluorescent dyes or ions conjugated to fluorescent tags; and (c) optical sectioning of thick sections of embedded specimens for three dimensional reconstruction of cellular and tissue elements. The Nikon PCM 2000 System will address the immediate needs of a diverse group of biomedical scientists whose research programs will benefit significantly by the availability of this much needed instrumentation.
