Services


Services

The Microscopy Shared Resource (MSR) offers training tailored to the needs of the individual investigator on all microscope systems in the facility. Unique to the facility, consultation and support on subjects ranging from experimental design, troubleshooting and data analysis are also offered. Processing and analysis software for image presentation and publication is also available to Ohio State cancer researchers.

The MSR is the only facility on campus that provides processing of biological samples for evaluation using electron microscopy. These services include sample processing of TEM and SEM samples requiring different treatments in order to obtain optimal results. For SEM sample processing, the researcher can choose between chemical dehydration or critical point drying depending on the application and desired result. Traditional epoxy resin processing and sectioning are available for TEM sample observation. However, examination of soft materials such as biopolymers used as substrates for cell growth is very difficult to section using this approach. For optimal imaging of soft samples, a cryo-ultramicrotome is available to section these samples after freezing. Cryo-ultramicrotomy is also offered for observation of samples using light microscopy to assist in eliminating out of focus light, an issue that is often a result of autofluorescence. The MSR also offers negative staining as a means of observing samples when dehydration is not suitable for maintaining structure. 

Instrumentation Equipment

The MSR offers the following instruments and equipment for use by members of the OSUCCC – James. 24-hour access to these instruments is offered to users who demonstrate the appropriate level of proficiency.

The FEI Tecnai Spirit BioTWIN Transmission Electron Microscope (TEM) is specifically designed for examination of biological specimens. The 120 kV TEM shows the fine structure of cellular organelles, details of cell-cell interactions and the distribution of extracellular matrix. It is also used for high- resolution examination of negatively stained nanostructures to determine their size and shape. 

The FEI Nova NanoSEM is a scanning electron microscope (SEM) equipped with a field emission gun and a through-the-lens that provides high-resolution digital imaging ideal for examining sample surfaces detectors. The ultrahigh resolution of up to one half million times magnification is used to examine structures including, but not limited to, cell surfaces, extracellular matrices, biofilms and nanostructures.

The Olympus FV 1000 Filter Confocal microscope offers routine confocal microscopy of fixed tissue sections and cultured cells. This system is equipped with four lasers, four fluorescence detectors, a DIC detector and a range of objective lenses including a 60x lens optimized for near UV imaging.

The Olympus FV 1000 Spectral Confocal system is also used for routine confocal microscopy of fixed tissue sections and cultured cells. Consistent with the Filter system, the Spectral system is equipped with four lasers, four fluorescence detectors, a DIC detector and a 60x objective lens corrected for near UV imaging. Two of the fluorescence detectors have spectral capabilities that enable the researcher to examine unique fluorophores and/or better eliminate autofluorescence. 

The Olympus FV1000 Multiphoton Laser Scanning Confocal System is equipped with a Deep See MaiTai TiSapphire laser, a 25x, 1.05 N.A water-immersion objective lens with a 2mm working distance, four non-descanned detectors and a forward, second harmonic generation (SHG) detector for collagen detection without the addition of a fluorescent marker. Infrared excitation using the two-photon system provides optical sectioning deeper than conventional confocal microscopy (up to 1mm). Because two-photon excitation occurs only at the focal plane, photobleaching is only confined to this area of the specimen. Consequently, this technique can be less phototoxic, making it ideal for live animal imaging. Additional support equipment for live animal imaging is also available, including a vital signs monitor and animal temperature control. This system also functions as a standard confocal instrument with four laser lines, three fluorescence detectors (two spectral) and a DIC detector.

The Visitech Infinity3 2D Array Confocal live cell imaging system has five lasers and a Hamamatsu C9100 EM13 CCD camera. The confocal scan head can be moved between an Olympus IX81 and an Olympus fixed stage microscope for imaging cultured cells or live animals. The confocal is ideally suited for imaging single color, rapid physiological events down to within 5 milliseconds at an imaging rate of 200 frames per second. 

The CRI Nuance Spectral Camera is available for use at the facility in conjunction with the Zeiss Axioskop microscope, or researchers may also use the Nuance camera with their own microscope system. Using tunable filters, the Nuance camera can acquire a 10nm spectrum at each pixel. The various signals from multiple labels in the resulting image are “unmixed” by the software, and a composite image is produced. This system enables researchers to separate labeling by color- specific signals in samples prepared for either bright-field or wide-field fluorescence microscopy.

The Zeiss Axioskop microscope is capable of capturing brightfield, darkfield, phase contrast and fluorescence images. Standard digital images are generated with a QICLICK-F-M-12 CCD camera (QImaging) equipped with an RGB liquid crystal color filter module for capturing color images. This camera can be moved to the stereo microscope.

The Olympus SZH Zoom Stereo Microscope is available for low magnification imaging (whole mount) and has a transmitted light illumination base with brightfield and darkfield illumination modes.

The Leica UC6 Cryo-Ultramicrotome is used for sectioning standard epoxy blocks or fixed frozen tissue for observation using transmission electron microscopy. This technique can also be used to obtain 0.2 micron thick sections for immunocytochemistry.

Two Reichert UltraCut E Ultramicrotomes are available for researchers to section resin embedded tissue samples.

The Leica CM1510S Cryostat enables the researcher to obtain 5-40 micron tissue sections for examination using light microscopy.

Fees

TEM   
$35 per hour
TEM sample preparation (per three samples) based on sample type

Tissue
$100

Cell Monolayer
$110

Cell Pellet
$120

TEM sample sectioning (Thick and Thin – per block)
$80

TEM sample sectioning (thick sections only – per block)
$40

SEM
$35 per hour 

SEM sample preparation (per three  samples of typical size)
$90

Single-Photon Confocal LSM (9-5)
$30 per hour

Single-Photon Confocal LSM (after-hurs)
$25 per hour

Multi-Photon Confocal LSM
$35 per hour

Visitech Infinity3 Live-Cell Confocal
$25 per hour

Light Microscope
$15 per hour

Ultramicrotome
$20 per hour

Cryo-Ultramicrotome
$30 per hour

Leica CM1510S Cryostat
$15 per hour

Vibratome
$20 per hour

Critical Point Dryer
$25 per run

Sputter Coater
$10 per run

Technical assistance (applies to all charges except sample preparation)
$35 per hour

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