Microscopy Facility

Equipment and Services                                

Widefield Microscopy - Conventional microscopy utilizing standard techniques in brightfield, darkfield, fluorescence, phase contrast and differential interference contrast is available on a variety of instruments in the main Core Facility room B78. Multiple upright and inverted microscopes capable of low to high magnification documentation are available with individual image capture workstations networked to the Microscopy  server.
Instrument Specifications

Live-Cell Time-Lapse Microscopy – Using the Nikon TE300 inverted microscope in room B78 with its ImagePro workstation, we can run time-lapse studies of cells grown in culture. With the new incubation chamber, XY stage and Z axis controller, we can track multiple areas in separate wells at the same time in multiple fluorescent channels. Our longest session to date was 6 days of uninterrupted imaging in Phase Contrast and 2 fluorescence channels. Cells may be grown in 6 well plates or 35mm dishes standard, but we can accommodate other vessels as needed.
Instrument Specifications

Low Magnification and Photomacrography
Specimens that are too large for a standard microscope require flexible equipment configurations for illumination and imaging conditions that vary depending on the specimen. We have the ability to image anything from whole mount slides to entire Petri dishes, as well as gross specimen
photography of excised biological tissues using our stereomicroscope or digital SLR-based macro equipment.

Confocal Microscopy - A Leica TCS SP2 confocal system is available for high resolution single cell observations, thick specimen analysis and co-localization studies. This system has 3 lasers and single wavelength emission spectra with the ability to carry out 3-D reconstruction and energy
transfer analysis. Sample preparation is generally done by the interested lab and analysis of the captured images can be done with the Leica analysis package or with 3rd party options available in the Core. FRET, FLIM, and FRAP capabilities are built into the software.
Instrument Specifications

2 Photon Microscopy – 2 photon microscopy continues where traditional confocal leaves off. With laser excitations in the longer wavelengths, 2P allows deeper imaging and less phototoxicity. This system was designed to be used with in vivo studies, such as imaging into inguinal lymph nodes,
while the subject animal is anesthetized, but it is also used with explanted specimens such as lymph nodes and brain slices, kept viable in heated media. Thick, fixed specimens, such as spheroids and skin reconstructs also benefit from the increased penetration and imaging capabilities of this system.
Instrument Specifications

Small Animal Whole Body Luminescence and Fluorescence Imaging
The Xenogen IVIS 200 whole body imager is designed for detection of weak fluorescent or bioluminescent signals deep in living mice for non-invasive longitudinal studies of growing (or regressing) tumors in mouse models. The system is housed in the Institute’s barrier mouse facility and is only
available to those researchers with approved protocols and animals housed in the colony. The integrated photon counting software can normalize the data for direct comparison to future studies or to similar instruments anywhere in the world.

Specialty Image Capture - The facility maintains equipment to provide “traditional” photographic support, such as photography of small animals (mice), experimental set-ups, copy stand specimens, or even people when needed, as well as special equipment for unusual situations such as GFP fluorescence in large specimens like adult mice. The Nikon D200 and Kodak 14n digital cameras have a variety of interchangeable lenses, depending on the situation, and special portable studio lighting is also available as needed.

2, 3, 4, and 5D Image Analysis
Capturing images is only the first step in scientific documentation. The images also need to be analyzed to extract meaningful data. The facility maintains multiple workstations and a variety of software packages for
this purpose with internal support and training from the staff, or custom support available from the suppliers for special cases.
Software Specifications

Microscopy Support Services:

• Training - includes Core equipment training, image processing and analysis, and digital imaging ethics. Training is mandatory prior to unassisted use of all Facility equipment.

• In-Lab Microscopy Assistance - The Microscopy Core staff is also available to assist researchers with their own equipment on site. All microscopy and imaging equipment is somewhat unique and often a minimal amount of instruction will help researchers realize better results from their existing set-up.

• Consulting – Consultation with investigators to determine specific imaging needs and to aid in designing experiments that make the best use of available equipment.