Core B will support the establishment and maintenance of the COBRE Confocal Microfluorometry and Microscopy Core facility. The Core will provide the instrumentation, expertise, training and mentoring in confocal microfluorometry and microscopy necessary for the advancement of all four COBRE projects. These projects depend critically on microfluorometry of fast Ca 2+ signals in cultured and native epithelial tissues, on the subcellular visualization of molecules including the 3-dimensional reconstruction of labeling, on coqocalization experiments, where maximal resolution and color separation through emission fingerprinting are necessary and on advanced co-localizations techniques such as FRET, for which emission scanning is critically. Based on our expertise and experience and with the goal to advance the mentored projects, we have identified the following components for this facility. A fully-motorized inverted microscope (Axiovert 200, Carl Zeiss), a confocal scan module with emission scanning capabilities (LSM 510 META, Carl Zeiss), a laser module with four lasers (Carl Zeiss), an electronics and computer module (Carl Zeiss), a power-line stabilizer, a vibration damped table (Technical Manufacturing Corp.), two temperabxre-controlled superfusion systems (at least one of them will in part be custom-made at the Physics Department Machine Shop at Kansas State University), two manipulators (MN151, Narishige) and a microinjection system (InjectMan with FemtoJet, Eppendorf). The superfusion systems will consist of a closed superfusion chamber specialized for the use of cultured epithelial cells grown on permeable support and of an open superfusion chamber for mounting native tissues with the help of two fine glass needles guided by the two micromanipulators. This instrumentation in conjunction with the provided expertise, training and mentoring will enable COBRE investigators to establish themselves through advancing their research on epithelia in health and disease.
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