Scientific Report 2003-2004 - Cleveland Clinic Lerner Research ...

IMAGING COREIMAGING COREDIRECTORJudith A. Drazba, Ph.D.HISTOLOGY MANAGERLinda VargoELECTRON MICROSCOPYMANAGERMei YinDIGITAL IMAGING TECHNOLOGISTSDmitry LeontievJoydeepSarkarPostdoctoral FellowAmit Vasanji, Ph.D.The Imaging Core Facility consists of three divisions: Digital Imaging, Histology, and ElectronMicroscopy. The facility provides a wide range of advanced imaging and scientific consultation servicesfor all investigators in the Lerner Research Institute.The primary mission of the facility is to assist investigators in producing high-resolution images ofcells and tissues using light and electron microscopy. For those requiring basic light microscopy the corewill help determine appropriate staining protocols and assist with the proper use of one of two newupright Leica DMR microscopes equipped for transmitted light and fluorescence microscopy. Thesemicroscopes are used for observing specimens mounted on slides. Digital images can be captured with ahigh-resolution Princeton Instruments MicroMax cooled CCD camera using ImagePro Plus Capture andAnalysis software. The microscopes are equipped with a full range of Chroma filters for fluorescentimaging. Living specimens in dishes or flasks can be observed on the new inverted Leica DMIRBfluorescence microscope. This microscope is also equipped with a PI MicroMax cooled CCD camera,ImagePro Plus Capture and Analysis software, and a full range of filters for standard fluorochromes andfluorescent protein tags. Another inverted Leica DMIRB fluorescence microscope is equipped for livetime-lapse imaging with a high-resolution Photometrics CoolSnap cooled CCD camera, a Sutter filterwheel, Uniblitz shutter, Prior Z-focus motor, stage-mounted heat/CO2 incubator and MetaMorphsoftware.Those requiring optical sectioning capability to better resolve the fluorescence in their samples canuse one of two new state-of-the-art Leica TCS-SP spectrophotometric laser scanning confocal microscopes.These instruments provide three-dimensional information from the sample and are eachequipped with four lasers for excitation at 351, 364, 457, 488, 514, 568 and 633 nm. Emitted light canbe detected interactively from 350-800 nm in 5-nm increments. We are now routinely imaging quadruple-labeledfluorescence specimens.The facility provides a full range of histology services including processing and embedding inparaffin, sectioning, H & E staining, a broad range of special stains, and frozen sectioning. Conventional(non-fluorescent) immunostaining of cells and tissues is also available.The EM services comprise conventional transmission electron microscopy, as well as antigenlocalization by immunogold labeling techniques. Conventional TEM services include routine processingof samples, (glutaraldehyde-osmium fixation, dehydration and plastic embedding), thick or ultra-thinsectioning, and photomicrography of thin sections. Possible samples include fresh tissue specimens,cultured cells growing on a substrate, cells in suspension, or subcellular fractions (for purity determinations).Sub-cellular localization of antigens within cells by immunogold localization can also be performed.In addition to the microscopy services investigators also have access to the Image Processing lab.This includes several NT workstations, a flatbed color scanner, a photographic quality Fujix Pictrographyprinter, and a high-resolution Polaroid Sprintscan 4000 slide scanner that can scan conventional 35 mmslides or specimens on microscope slides that are too large for conventional compound microscopy. Thisequipment provides investigators with sophisticated tools and assistance for preparing their images forpresentations and publication.This year, the facility has acquired a Leica MicroDissection microscope that will allow investigatorsto precisely excise cell groups, single cells or even parts of cells out of tissue sections or cultureswithout touching them or contaminating them. Such explants could then be used for RNA, DNA orprotein analysis.Web site: http://www.lerner.ccf.org/services/imaging/184