SCIENTIFIC REPORT 2004 - Sylvester Comprehensive Cancer Center

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S H A R E D R E S O U R C E S S H A R E D R E S O U R C E S A N A L Y T I C A L I M A G I N G C O R E MANAGER Alberto Pugliese, M.D. Associate Professor of Medicine CO-MANAGER Beata R. Frydel, Ph.D. Associate Scientist of Neurosurgery PURPOSE Many research endeavors rely on state-of-theart imaging and molecular histology techniques requiring the use of complex and costly equipment not practical for the individual investigator to acquire and maintain. The Analytical Imaging Core is a campus-wide resource spearheaded by the Diabetes Research Institute and UM/Sylvester, with seed support from the dean of the University of Miami School of Medicine. The Juvenile Diabetes Research Foundation awarded additional support for a five-year period in December 2003. The core’s main goals are to: • Provide access to sophisticated, modern instrumentation for imaging and molecular analysis of tissue and cellular specimens to investigators. • Provide expertise, guidance, and training to core users and help them optimize protocols for their applications that can be shared with other investigators. SERVICES This core provides the following services: 1) Confocal Microscopy: Confocal microscopy offers many advantages over standard fluorescent microscopy including increased sensitivity, resolution, and the ability to image relatively thick, fluorescently labeled biological specimens in two or three dimensions. Confocal microscopy creates an “optical section” of the cells or tissues being imaged and an increase in effective resolution due to a large increase in signal-to-noise ratio. As a result, outstanding images can be collected from cells and tissue sections that would otherwise yield little or no information. The workhorse instrument for confocal microscopy is the Zeiss LSM-510, which can detect up to five channels and four fluorescent channels simultaneously—from UV to far red, plus a separate detector for transmitted light. The outstanding beam control afforded by the Zeiss LSM-510 makes it an ideal instrument for other advanced fluorescent applications such as fluorescence resonance energy transfer (FRET), fluorescence recovery after photo bleaching (FRAP), or ratio-imaging for fluorescence quantitation. The core also is equipped with an Atto Instruments spinning disk confocal microscope (CARV), a confocal instrument particularly suited for live cell analysis, and video-rate (30 frames per second) imaging. 2) Standard Epifluorescence Microscopy: The core also is equipped with a Leica DMIRB inverted microscope capable of performing triple fluorescence, phase contrast, and light microscopy, etc. UM/Sylvester Comprehensive Cancer Center Scientific Report 2004 143
S H A R E D R E S O U R C E S 3) Laser Scanning Cytometer (LSC): The LSC allows “flow cytometer-like” fluorescent imaging and quantitation of tissue sections on a microscope slide. The LSC records the position and time of measurement for each cell analyzed so that multiple biochemical, immunological, and morphological measurements can be made on each cell. Possible applications for the LSC include: detection and quantitation of apoptosis (TUNEL, annexin); in situ hybridization (FISH); and the study of cell adhesion, cell cycle, and DNA content, etc. 4) MetaMorph Imaging System (MIS): This imaging system consists of hardware and software that enables the capture and analysis of microscopy and digital images obtained using the instruments described above. 5) Laser Capture Dissection Microscope (LCM): The LCM can dissect portions of tissues (or even single cells) from cell smears and fixed and frozen tissue sections, obtaining essentially pure samples of a desired cell population (500- 1,000 cells per hour). The dissected cells can then be used to extract RNA, DNA, or proteins for further studies. LCM offers unprecedented access to specific cells for defining their pattern of gene expression, in combination with powerful techniques such as gene array and real-time PCR. This technology is particularly powerful in the study of human diseases and several cancer applications, where only small amounts of tissue may be available for study. 144 UM/Sylvester Comprehensive Cancer Center Scientific Report 2004
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SCIENTIFIC REPORT 2004 - Sylvester Comprehensive Cancer Center

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