The Confocal Shared Laser Scanning Microscope Facility (CLSMF) has been active on the UNMC campus since 1996 when it was established through institutional funding. The objectives of the CLSMF are as follows: 1. To fill the need for high resolution image analysis of both live and fixed tissue and cultured cells. 2. To provide training in the use of analysis tools for live cell component analysis of FRET, FRAP and FLIP experiments. 3. To provide essential hard and software for analysis of collected data. 4. To provide the expertise needed both in preparing materials for microscopic examination and in carrying out image generation and analysis The CLSMF is located on the first floor of the Durham Research Center (DRC), Room 1056. The facility contains two Zeiss Confocal Laser Scanning Microscopes. One is a fully motorized LSM 510 META equipped with an Argon Laser (458/477/448/514 nm), a DPSS 561 nm and a HeNe 633nm as well as Nomarski optics. This allows superimposition of signals from fluorphores to be overlain on a DlC image of the cells. Zeiss software is available for collection and analysis of FRET, FRAP and FLIP data, reconstruction of 3-D images, Z-sections, time series, etc. Images can be stored directly on CDs, DVDs, etc. or sent directly to Biomedical Communications by network transfer for production of high resolution dye sublimation prints, slides, posters and illustrations. The Zeis Axiovert inverted microscope used with this system has lOx, 20x, 40x oil, 62x oil and 40x water objectives and a Z-axis motor as well as an xy stage motor. All functions are computer automated. In addition, controlled environment chambers (moisture, C02 levels, temperature) are available for studies of live cells. Judith K. Christman, Ph.D. Is the Facility Director. She has extensive experience in developing immunohistochemistry protocols and In analyzing confocal images. These skills enable her to advise users as to how to optimize their use of confocal microscopy for a variety of different cells and tissues.
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