This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In FY2006, the Imaging Core maintained the growth in confocal microscopy utilization that was described for FY2005. A major advancement in the capabilities of the Core in FY2006 derived from the acquisition of the Olympus FluoView 1000 confocal microscope which is currently located in the facilities of the Cardiovascular Research Institute. This new instrument offers all the functions of the FVX microscope that was originally included within the COBRE grant. The FV1000 automates these functions with computer control over the microscope and confocal acquisition parameters. The FV1000 allows acquisition from three fluorescence channels, permitting an assessment of the co-localization of three proteins within a cellular or tissue sample. These three fluorescence channels can be assigned to a wide variety of cellular dyes with excitation wavelengths in a range from 405nm (e.g. the nuclear stain DAPI) to 675nm (Cy5). This broad range of dye options offers investigators the opportunity to utilize the fluoro-chrome that provides the highest level of sensitivity and selectivity for their studies. This level of flexibility is permitted by the precise definition of excitation and emission wavelengths that is provided by the FV1000. The Core also installed a microscope mounted, physiologic chamber that will permit the acquisition of confocal images from living cells subjected to a variety of experimental conditions over a series of time points. These experiments can include fluorescent substrates that are micro-injected into viable cells. The cellular localization of enzyme activation can be determined in vivo by the appearance of fluorescence subsequent to enzyme activation.
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