The Center for Microscopy and Image Analysis (CMIA) at The George Washington University - Medical Center (GWUMC) provides core resources for acquisition and processing of microscopic images of relevance to biomedical sciences. The CMIA was made an institutional core facility in 1999 by a commitment of considerable institutional support. It has remained true to its original goals of research, dissemination of knowledge, and educational opportunities at several levels. CMIA offers outdated BioRad MRC 1024 ES confocal laser scanning microscopes, the maintenance of which is seriously challenged by the lack of service contracts. We now seek to acquire the next generation of confocal laser scanning microscope (CLSM) - Zeiss LSM 710. The requested CLSM will be equipped with variety laser excitation protocols and spectral emission detection, and environmental control to address current equipment deficiencies and to meet the research objectives of NIH funded investigators. In addition to the significantly higher detection efficiency and reliability compared to the existing CLSM, the proposed instrument will be capable of laser stimulation, allowing the implementation of sophisticated visualization methods such as fluorescent resonance energy transfer (FRET), photobleaching and photoconversion. Spectral detection will provide the researchers with a higher level of flexibility in selecting the emission bandwidth, elimination of cross-talk and autofluorescence. Additional laser lines (405nm) will offer labeling detection for state of the art markers such as quantum dots. Proposed instrument will be equipped with environmental control, which in combination of the greatest flexibility offered by excitation and emission protocols will advance investigators life cell imaging experiments - not possible with the current instrument. The CLSM will be the first of its class in the institution and will be capable of simultaneously detecting spectral signatures from up to eight fluorophores even if they have overlapping emission spectra, by using linear un-mixing algorithms. Proposed CLSM will advance fluorescence labeling detection, will provide us with the capability for studying protein-protein interactions, and will allow us for applying new and more reliable labeling strategies. A group of well established NIH funded investigators support this application. These investigators will be able to advance their projects beyond the limits imposed by the existing BioRad 1024ES CLSM. CMIA will use its multi-user environment for publicizing the achievements of investigators with the proposed CLSM, which will create a broad platform for methodological sharing and advancement of all NIH funded investigators. Thus, the instrument will foster interdepartmental interactions and collaborations. Training with the instrument will be available to our Institute of Biomedical Sciences PhD program students on research rotations and dissertation projects.
The deficits imposed by the available confocal laser scanning microscope available at The George Washington University Medical Center core facility are addressed in this application. Acquisition of a state of the art confocal laser scanning microscope is proposed, which will be integrated in into the core facility. The new instrument will allow a group of well established investigators to advance their NIH funded project.
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