This is an application for a shared instrumentation grant to establish a Laser Scanning Confocal Microscope (LSCM) facility at the Tufts University Schools of Medicine. The BIORAD MRC-600 LSCM system, will extend the research capabilities of five investigators, and their groups, by providing contemporary instrumentation with which to analyze living and fixed cells and tissues labeled with a range of fluorescent probes. A major user group of five NIH-funded scientists will share access to a LSCM uniquely configured to optimize the study of whole tissues, thick sections, and isolated multicellular aggregates derived from brain, liver, ovary, and their associated vascular elements. The LSCM configuration incorporates: (a) inverted light optics for specimen manipulating, (b) multiple excitation capability through a Krypton/Argon ion mixed gas laser illumination source (c) three dimensional reconstruction software and image archiving capabilities and (d) data transfer capabilities for transporting confocal image data sets to preexisting image analysis workstations. Specific applications that this instrumentation will provide include: (a) detection and spatial discrimination of reporter fluors in single, double or triple labeled samples; (b) monitoring of intracellular activities with several vital fluorescent dyes; and (c) optical sectioning of thick fixed specimens for 3D reconstruction of cells and their constituents in intact tissues. The requested instrumentation will benefit the immediate needs of a diverse group of biomedical scientists whose research programs continue to rely heavily on the availability of light optical imaging methods.
