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Multi-photon Confocal Microscope

The Center's Imaging Facility is equipped with a Zeiss LSM 510 NLO META system for state-of-the-art confocal microscopy, using a Zeiss Axiovert 100M inverted microscope. The illumination system incorporates one argon and two helium-neon lasers for single-photon excitation at 458, 488, 514, 543, and 633 nm, and a Coherent tunable titanium-sapphire laser for two-photon excitation (700-900 nm). The META spectral imaging system plus two conventional photomultiplier detectors offer simultaneous multi-channel color display of fluorescence. The 32-channel META spectral detector allows the separation of signals with extremely overlapping emission profiles, heretofore impossible using glass filters. For example, GFP and FITC may be separated with confidence. A transmitted light detector permits recording of brightfield and DIC images. User-friendly software features include 3D reconstruction, animation, time series, user-definable regions of interest for photobleaching and uncaging studies, multi-tracking to eliminate bleed through, emission fingerprinting, linear unmixing , quantification of colocalization , scale bar overlay, and a reuse button to duplicate system settings from a previously acquired image.

Two-photon scanning microscopy represents a powerful tool for investigating living cells and very thick specimens. Due to the non-linear characteristics of two-photon excitation, effective excitation takes place only in the focal spot of the objective lens. The highly localized excitation results in reduced photobleaching and phototoxicity, a distinct advantage for physiology experiments, including techniques like FRAP (fluorescence recovery after photobleaching ) and FRET (fluorescence resonant energy transfer). Live cell work is also facilitated by a temperature-controlled microincubator perfusion chamber (Harvard Apparatus), which may be mounted on the microscope stage.

Investigators interested in using the Confocal microscope can contact Howard Rees, PhD at 404/727-8043 or hrees@emory.edu.

Light Microscopy

Workstations are available in Whitehead Room 525 for capturing, analyzing, and printing digital images. Specifically, the five workstations are equipped as follows:

1. Power Mac 8500 computer, Olympus BX60 microscope for brightfield and DIC, and Kodak EOS-DCS5 color camera.

2. Power Mac G3 computer with Stereologer software, Nikon Microphot -SA microscope, Sony color camera.

3. Dell computer with Zeiss LSM510 Image Examiner software for confocal images and .

4. Dell computer with MetaVue image analysis software, Hamamatsu monochrome camera, Zeiss Axiovert inverted microscope for fluorescence and phase contrast, Leica DMRBE upright microscope for brightfield and fluorescence.

5. Dell computer with MetaMorph image analysis software, Leica DC500 color CCD camera, Leica M420 macroscope for low-power work, Northern Light illuminator.

Investigators interested in using the Imaging Facility can contact Howard Rees, PhD at 404/727-8043 or hrees@emory.edu .

Electron Microscope Imaging Facility

The Emory Neurology Microscopy Core Laboratory at Woodruff Memorial Research Building provides campus-wide service and training in biological transmission electron microscopy. In 1997, the facility acquired a state-of-art Hitachi H -7500 transmission electron microscope through an NIH Shared Instrumentation Grant. The facility is also equipped with all the necessary instruments for EM sample preparation, data processing and analysis. These include two Leica UltraCut S ultramicrotomes for thin sectioning, Leica AFS automated freeze-substitution, a Gatan Bioscan CCD camera for digital image acquisition on TEM, and both conventional and digital photographic darkrooms. In addition to conventional electron microscopy, ENMCL is highly regarded nationally for its strength in immunoelectron microscopy. The staff at the ENMCL has many years of experience and have published extensively addressing various techniques and applications of immunoelectron microscopy.

Investigators interested in using ENMCL's services can contact Hong Yi at (404) 712-8491 or hyi@emory.edu

Laser Capture Microdissection

A Pixcell II laser capture microdissection system is available for collecting single cells, cell layers, or groups of cells from tissue sections or cell cultures. The procedure allows the investigator to visually select cells of interest for subsequent molecular analyses. The system is equipped with an inverted microscope with bright-field and fluorescence optics and a non-destructive infrared laser for dissecting the tissue.

Investigators interested in using the Laser Capture Microdissection microscope should contact Michael Iuvone , PhD at 404-727-5989 or miuvone@pharm.emory.edu.