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Microscopy

Equipment

Learn about the equipment provided by the Microscopy Shared Resource (MSR) to OSUCCC – James cancer researchers.

    Facility Online Manager (FOM)

    The MSR offers the following instruments and equipment for use by OSUCCC investigators. Twenty-four-hour access to these instruments is available to trained individuals who demonstrate the appropriate level of proficiency.

    The MSR pools resources from two premier imaging sites at The Ohio State University: the Campus Microscopy & Imaging Facility (CMIF) and the Center for Electron Microscopy and Analysis (CEMAS).

    OSU Comprehensive Cancer Center Microscopy Shared Resource (MSR) Letters of Support for a new TEM

    Instrumentation

    Confocal Microscopy

    The MSR offers standard laser-scanning confocal microscopy for examination of fixed tissues or cells in cultures at the highest possible resolution. 

    • The two inverted Olympus FV 3000 confocal microscopes enable visualization of slides, petri dishes and well plates, and enable tiled imaging to produce high resolution mosaics. One FV3000 is equipped with five lasers covering a wide range from 405 to 750 nm, a wide range of objectives and five detectors, including two very high sensitivity detectors capable of visualizing the weakest fluorescent signals. The second FV3000 is equipped with 7 lasers covering the range from 405 to 647 including laser to separate CFP and YFP, a wide range of objectives and four detectors, including two very high sensitivity detectors capable of visualizing the weakest fluorescent signals.
    • The two upright Olympus FV1000 confocal microscopes are equipped with four lasers and four fluorescence detectors, as well as a DIC detector for imaging non-fluorescently labeled cells and tissues. Ohio State researchers receive a combination of classroom and hands-on training to prepare them for full use of the confocal by their third training session.

    Cryo-Electron Microscopy (Cryo-EM)

    The MSR offers access to a comprehensive suite of instruments needed for cryo-electron microscopy (cryo-EM). This powerful technique allows for the observation of biological specimens in their native environment by embedding samples in vitreous ice. Cryo-EM has transformed structural biology, and its use is expanding into other fields, such as small molecule crystallography, infectious disease, and cell biology.

    • The Thermo Scientific Glacios is used for sample screening and subsequent single-particle data collection. The instrument can also be configured for microcrystal electron diffraction (MicroED). The Glacios is equipped with a fully integrated Falcon 3EC direct electron detector, which provides high sensitivity and makes this instrument a standalone single particle analysis (SPA) data-acquisition solution capable of generating density maps at resolutions of 4 Å and below.
    • The Thermo Scientific G3i Krios provides near-atomic resolution imaging of molecular structures via SPA and nanometer resolution of cellular features using cryo-tomography. The Krios includes a Gatan BioQuantum K3 imaging filter coupled with the K3 direct-detection camera, as well as a phase plate to further improve contrast. Both cryo-TEMs have a cryogenic sample manipulation robot (autoloader) that facilitates the direct exchange of samples between the Glacios and Krios.

    Focused Ion Beam Microscopy (FIB)

    Light Microscopy

    The MSR provides instrumentation for producing high-quality, light-level, digital images for Ohio State investigators. The wide-field microscope is equipped with a high-sensitivity CCD camera with bright-field, phase-contrast and fluorescence capabilities. High sensitivity wide-field microscopy can image extremely dim samples, which gives this technique an advantage when using fluorescence. The camera can also be used with the Olympus SZH Zoom Stereo Microscope system for low-magnification imaging of whole-mount or large samples not suitable for the wide-field system. The stereo microscope has an auxiliary fluorescence light source called the NIGHTSEA viewing system that is capable of illuminating samples containing green fluorescence markers (e.g., GFP).

    Live Cell Imaging 

    The MSR provides access to cutting-edge instrumentation that can image the dynamic processes of cancer formation, growth and metastasis. 

    • The Nikon A1R live cell imaging system features a resonant scanner for rapid confocal imaging with four fluorescence detectors or a spectral detector, high-speed image capture for calcium fluctuation analysis, multi-positional imaging of live cells, mosaic or tiled imaging resulting in images much larger than the standard field of view, and high-throughput imaging and analysis capabilities. The Nikon A1R confocal microscope has four lasers, four single channel fluorescence detectors, a 32-channel spectral detector and a transmitted light detector for bright-field and DIC imaging of both fixed and live cells. High-throughput imaging is particularly useful for experiments performed in 96- or 384-well plates, as well as for drug discovery assays wherein researchers can rapidly screen the effects of various drugs or concentrations on live or fixed cells. The Nikon A1R is also equipped with a full microscope incubation enclosure that, when paired with the Perfect Focus System, allows for drift-free time-lapse imaging as short as milliseconds or as long as days. The system is housed in a BSL2-compliant space that facilitates live-cell imaging of a broad range of specimens.
    • The Nikon AXR live cell imaging system features a resonant scanner for rapid confocal imaging with four fluorescence detectors or a spectral detector, high-speed image capture for calcium fluctuation analysis, multi-positional imaging of live cells, mosaic or tiled imaging resulting in images much larger than the standard field of view, and high-throughput imaging and analysis capabilities. The Nikon AXR has faster scanning capabilities for quicker imaging and reduced photobleaching. In addition, it has an expanded field of view, capable of up to 8K x 8K resolution for the Galvano scanner and up to 2K x 2K resolution for the resonant scanner. The Nikon AXR is equipped with a stage top incubator that, when paired with the Perfect Focus System, allows for drift-free time-lapse imaging as short as milliseconds or as long as days. The Nikon AXR confocal microscope is housed in a BSL2-compliant space in the Pelotonia Research Center.

    Multiphoton Imaging

    The Olympus FV1000 multiphoton microscope (MPE) is mounted on an upright stand and is highly specialized for live-animal microscopy using rats and mice. The MPE can penetrate tissues up to 1,000 um with an infrared laser (IR), providing a significant advantage over standard single-photon confocal microscopes (maximum penetration of 30 to 50 um). The MPE is equipped with a Spectra Physics DeepSee, TiSaphire IR laser and an IR-corrected 25X/1.05 numerical aperture objective lens with a 2 mm working distance. This combination of deep tissue penetration and live-animal imaging gives the MPE potential for the intravital staining for tracking cells in live animal tissues. The system is housed in a BSL2-compliant space that facilitates live-ell imaging of a broad range of specimens.

    Scanning Electron Microscopy (SEM)

    Using SEM, researchers can examine samples without the traditional metal coating. OSUCCC researchers use SEM to visualize tumor microenvironment and nanostructures, especially for the development of cancer drug delivery systems. Observing the three-dimensional surface relationships between cells and the tumor microenvironment can help researchers understand cellular growth patterns and how anticancer treatments affect cells.

    Super Resolution Microscopy

    The Nikon Super Resolution Microscope is capable of performing Structured Illumination Microscopy (SIM), STochastic Optical Reconstruction Microscopy (STORM) as well as Total Internal Reflection Fluorescence (TIRF) microscopy. The system can be used for live cell super-resolution imaging for sub-diffraction limit imaging with SIM or for single molecule imaging with TIRF. The super-resolution microscopy techniques enable visualization of extremely fine details. The STORM system enables researchers to visualize molecules within fixed cells down to a resolution of 20 nm. The microscope is equipped with the Perfect Focus System, which allows for drift-free time-lapse imaging as short as milliseconds or as long as days. This new capability will enable Ohio State researchers to explore the fundamental mechanics of living processes in real-time to rapidly facilitate the progress of biomedical research. The system is housed in a BSL2 compliant space facilitating live cell imaging of a broad range of specimens.

    Transmission Electron Microscopy (TEM)

    TEM examination of ultra-thin sections supports research of cellular organelles, cell-cell interactions, viral particles, and nanostructures proposed for chemotherapeutic drug delivery. TEM can be used to determine the subcellular distribution of a protein, using immunocytochemistry with gold-labeled antibodies.