Funds are requested for the acquisition of a modern Laser Scanning Confocal Microscope system to be installed within the Division of Core Research Laboratories at Rhode Island Hospital (the State's largest Brown University-affiliated teaching hospital and trauma center). The Nikon C1 Si Spectral Imaging Confocal Microscope will significantly benefit the research and teaching endeavors of the seven NIH-funded investigators whose projects are detailed in this submission as well as that of Brown University campus-based investigator, and the work of researchers at the five other Brown-affiliated hospitals which collectively comprise the Brown University Medical School. Also noteworthy among the efforts that will be supported by this instrument are four newly-established Centers of Biomedical Research Excellence (COBRE) within the local scientific community. We currently have a decrepit Nikon PCM 2000 Confocal Microscope, which has served us well for eight years. However, this is an outmoded system (in terms of hardware and software) that no longer functions properly. Our laser strength is down to approximately half-power and there are optical/image faults that the service engineers have not been able to correct. Moreover, the overall design and out-of-date software make it impossible to meet the needs of the research community we serve, especially as related to overall fluorescence sensitivity and with regard to live cell imaging. The requested Nikon C1 Si Spectral Imaging Confocal Microscope is a novel design that has only become available within the last year. It is a modular confocal system consisting of a TE200-E2 Motorized Microscope quipped with the following components: 1) a separate scan head; 3) a spectral detector module that allows simultaneous spectral data acquisition across 32 separate channels using a 32 PMT linear array . Moreover, the 32 PMT linear array allows 512 spectral image acquisition in 2 seconds, a capability that is unique to this instrument. The spectral resolution can be modulated by choosing channel widths of 10nm, 5 nm, and 2.5 nm over a range from 400nm to 750 nm: 4) a laser module which increases sensitivity by insuring that there is no down time while digitizers send data and reset; 5) separate control module, and monitor, thereby minimizing electrical interference or vibration from one component to another. Acquisition modes include confocal fluorescence, confocal spectral imaging, reflection imaging, interference reflection contrast, and scanned transmitted light in brightfield, phase contrast, and DIC. The C1Si is suitable for a wide variety of experimental analyses including: multiple color experiments; spectroscopy of environmentally sensitive probes like pH sensitive GFP; single pass FRET; removal of unwanted fluorescence by spectral analysis of dyes in-situ. All of the above can be done in motorized Z stack, time lapse mode. Corrections to this grant are so extensive that demarcations are impractical. Relevance: The requested instrumentation will not only support the transitional and basic science research of the seven Major User Group (MUG) projects described in this proposal, it will also support investigators of several Centers Of Biomedical Research Excellence (COBRE) including Rhode Island Hospital's COBRE For Cancer Research Development (the PI of which is a member of our MUG) as well as investigators of Brown University's COBRE For Cancer Signaling Networks, the Women and Infants' Hospital COBRE For Perinatal Biology, and the Roger Williams Medical Center COBRE for New Stem Cell Biology. The instrumentation will also potentially benefit investigators from Brown University and the six Brown University-affiliated teaching hospitals that are involved in biomedical research but are not part of the above COBRE programs. Finally, access will be provided to investigators from other colleges/hospitals, and universities in our area as needed. ?
