Eight Major Users request funds to establish a central digital deconvolution imaging station and two satellite facilities. The primary need is to acquire quantitatively accurate confocal images, but avoid the deleterious effects of laser illumination particularly photobleaching and in living tissues phototoxicity. The main station will comprise an epi-fluorescence microscope, two interchangeable cooled CCD cameras, a computer-controlled stage (X,Y and Z axes), a stage incubator for living cells, a Silicon Graphics R8000 computer for image deconvolution, and a color printer. In addition, we will establish two satellite facilities at the Health Sciences Center. These will be capable of routine image acquisition, and will be connected to the main station via ethernet, so that images can be deconvolved at the central station. The satellites will be useful for work that does not require the special capabilities of the central station. Each will comprise a microscope, Z-axis stepper motor, camera, computer, and ethernet link. In this way, we will be able to maximize the use of this technology. All eight Major Users receive NIH funding for their work, and all of the proposed projects arise from currently funded projects. About half of the Major Users have had extensive experience with digital image processing and/or laser scanning confocal microscopy. No comparable instrument is currently available at the Health Sciences Center. We have an eight year history of collegial implementation and management of previous Shared Instrumentation Grants a background that augers well for the success of the current proposal.
