Progress in cutting-edge biomedical studies has become increasingly dependent upon the technology required to measure biological process as they occur in living organisms, tissues, and individual cells. In recent years, increasing numbers of probes have been developed that can detect cellular process in live cells as have the technologies and tools to monitor these activities. Research groups at The Scripps Research Institute have initiated activities to image processed in live cells in several areas. Using green fluorescent protein (GFP) to track intra- and intercellular dynamics of many proteins, including movement of centromeric proteins during mitosis, targeting of proteins to the nuclear envelope, virus movement, neuronal cytoskeleton dynamics and monitoring expression of clock proteins. Others are developing and applying novel fluorescent analogs to indicators for studies in live cell function. Our goal is to establish at TSRI a unique facility with equipment and software that will enable us to study live cells in real-time with the capacity to capture light in optical sections, to deconvolve the capture light, carry out ratio image, and to reconstruct patterns of fluorescence through cells. The system will perform bright field and differential interference contrast as well as low-light fluorescence and luminescence imaging of animal and plant cells and tissues. We propose to acquire a Deltavision digital microscopy system that will emphasize the following features: optimized optical paths to reduce light loss, several types of stage chambers for the maintenance of living specimens during image acquisition, a high resolution/high readout rate cooled CCD camera, optical sectioning capabilities and proven deconvolution software for assembling 3D images, and silicon graphics workstations for rapid convenient image acquisition and processing. While each of the participants has existing fluorescence microscopy capabilities within their labs, this equipment will enable areas of research far beyond those currently available at this Institute. Furthermore, this Digital Microscopy Resource system for live cell processes will be included within the physical facility that also contains confocal and electron microscopes, and will serve a large diverse biology community at TSRI. It will also initiate opportunities for interdisciplinary collaboration with other well established TSRI programs in computational biology, imaging science, and biological structure through member participation on the Advisory board that will oversee the use of the instrument.
