Imaging Core (Digital Imaging, Histology, Electron Microscopy)
Cleveland Clinic Lerner Research Institute, NB1-55 Main Lab 1st floor
Judith A. Drazba, PhD, Facility Director
Contact:
| Linda Vargo, Histology Facility Support |
 216-444-5805 |
|
| MeiYin, EM Facility Support |
216-444-5805 |
|
| Digital & Confocal |
216-444-3760 |
|
|
|
|
Serves: All SOM Facility
Overview
Three divisions make up the Imaging Core at the Research Institute: Digital Imaging The Imaging core provides digital imaging, confocal and fluorescence microscopy, and laser microdissection. The core has three Leica DMR fluorescence microscopes equipped with Q-Imaging Retiga EXi cooled CCD cameras and Image ProPlus Capture and Analysis software for transmitted light (i.e. brightfield, phase contrast, differential interference contrast, darkfield) or fluorescence imaging. Two of the microscopes are designed for capturing images from fixed specimens on slides. The third microscope is inverted and can be used for short-term observation and imaging of live cells in flasks or culture dishes. For live time-lapse imaging over hours to days we have an inverted Leica DMIRB microscope equipped for both transmitted and fluorescence imaging with a heated stage and Leica environmental chamber, computer-controlled Sutter Instruments Lambda-10 motorized optical filter wheel, Roper CoolSnap HQ cooled CCD camera and Universal Imaging's Metamorph software. All microscopes are equipped with a full range of Chroma filters for fluorescent protein imaging. For confocal microscopy, the Imaging core has two uniquely capable Leica TCS-SP spectral laser scanning confocal microscopes each equipped with four lasers for fluorescence excitation at up to eight separate wavelengths. This allows us to visualize multi-labeled components of a specimen simultaneously to compare the three-dimensional relationships between them. For example we can visualize fluorescently labeled proteins and compare the quantity and localization relative to other labeled cellular components. For post-processing and analysis of confocal data we have several Dell Workstations and a Mac G5 running Improvision's Volocity software. These systems allow for the reconstruction of confocal optical slices into three-dimensional images using a variety of rendering methods. They also allow us to analyze and quantitate, three-dimensionally, the morphology and distribution of various labeled structures. The core also has a wide range of software, scanners, and printers for optimal image manipulation. Histology The histology services are administered as part of the Imaging core. A Ventana automated processor is used to dehydrate and infiltrate tissues with paraffin followed by embedding on a Sakura embedding station. Thin sectioning is then done on a Leica microtome. Routine H&E staining is available as are many special stains. Frozen sections can be provided using a Leica cryostat. Complete immunohistochemistry services are available using conventional (HRP- or Alkaline Phosphatase-tagged) labeling. The investigator provides the primary antibody and the core provides all secondary reagents including a variety of antigen retrieval techniques. Electron Microscopy The electron microscopy section of the Imaging core provides multiple services including embedding, thin and thick sectioning, pre and post-embedding immunoEM, sample observation in our FEI CM12 transmission electron microscope, and photomicrography. The core also maintains and trains users on the JEOL Scanning electron microscope, which is additionally equipped with elemental analysis. Users can get help fixing, dehydrating, critical point drying and sputter coating their samples in preparation for viewing and imaging in the SEM.
Page last updated: October, 2007
 |
