Cell Imaging Services
This facility is designed to provide state-of-the-art, quality-assured, morphologically-oriented services to investigators. It houses sophisticated microscopic equipment that provide digital images that can be subsequently processed and analyzed by available computer-based methods.
The Cell Imaging Facility Core provides the following services and equipment to projects directed toward elucidation of determinants of carcinogenesis. The facility is staffed by experienced laboratory technologists and research professionals who can provide guidance with experimental design, tissue sectioning, sample preparation, data analysis and interpretation, and guidance as to optimal or appropriate use of the instrumentation available in the facility.
- Morphologic services and consultation at the light and electron microscopy level
- Morphometry (semi-quantitative morphology)
- Light and electron microscopic immunocytochemistry
- Confocal scanning laser microscopy
- Laser scanning cytometry
- Atomic Force microscopy
- Scanning electron microscopy
- Laser capture microdissection
- Computer-assisted digital imaging and analysis
Equipment
The following equipment is available in the Cell Imaging Facility:
- JEOL 1210 transmission electron microscope with 32 bit internal microprocessor for instrument control; a Kodak MegaPlus high resolution video camera for digital image acquisition; and advanced image processing software (Image ProPlus). A Princeton Gamma Tech IMIX Microanalysis System is attached to this JEOL electron microscope for elemental analysis of specimens.
- JEOL T-300 scanning electron microscope for investigations of bulk specimens. Images can be captured in analog mode with an attached Polaroid camera, or can be digitally acquired via an RS-170 connection from the video output of the microscope to a video frame grabber in the Sun SPARCstation 5 workstation.
- Olympus BX50 research microscope for transmitted light, phase contrast, and epi-fluorescence microscopy. This microscope contains objective lenses of 4X, 10X, 20X, 40X and 100X , and bandpass filters for ultraviolet, blue and green fluorescence emissions. These filters allow single, or simultaneous double and triple fluorescence imaging. A PM30 automatic camera system is attached for conventional analog photography. A Low-light package for fluorescence microscopy is also attached to this microscope and consists of an Optronics MagnaFire digital camera. True color or grayscale digital images can be captured from this camera system onto a video frame grabber board in a workstation running Universe Imaging MeaMorph software.
- BioRad MRC 1024ES confocal scanning laser microscope system. This confocal imaging system was upgraded last year from the MRC 1000 model. The new system is a "true" 24-bit imaging system, providing enhanced pixel resolution over the previous model. This system consists of a scanning transmission assembly mounted on an upright Olympus BX50 light microscope; a 15 mWatt krypton/argon mixed gas laser delivering excitation lines of 488 nm, 568 nm, and 647 nm, allowing simultaneous visualization of up to three fluorophores; a polarizer kit for confocal reflection microscopy; a three-color transmitted light detector; BioRad LaserSharp software for quantitative evaluation of images; Confocal Assistant software for image processing and analysis; MicroVoxel (Indec Systems) software for 3-D reconstruction and analysis of confocal images; a Zip drive and cd recording drive for image storage, and an Epson Stylus Color 850 printer for working prints.
- Digital Instruments BioScope Nanoscope IIIa atomic force microscope station. This system consists of a BioScope atomic force microscope mounted on an Olympus IX70 inverted light microscope. Included software allows detailed image analysis and processing of digitally-captured images.
- CompuCyte Laser Scanning Cytometer. This system is a hybrid instrument between a flow cytometer and a microscope-based imaging system. It consists of an Olympus BX50 upright light microscope attached to a laser scanning system. The system is equipped with two lasers: an argon ion (approximately 488 nm) and a helium/neon (approximately 633 nm), and three filter block/photomultiplier tube combinations allowing for simultaneous imaging of up to three fluorophores, or two fluorophores and forward scatter. Software for analysis and data processing are part of the system, as well as a color ccd camera for image capture. Data and images are stored on 100 megabyte zip disks, and color working prints made with an attached Hewlett-Packard Deskjet 1600C printer.
- Zeiss Axioscope 2 wide-field fluorescence microscopy system. This system is fully equipped for fluorescence microscopy and digital image capture with a Hamamatsu digital ccd camera interfaced into a MacIntosh computer running Open Lab software.
- Arcturus PixCell II Laser Capture Microdissector system. This system is used for specifically removing desired cells from a tissue section. A cap containing a thermoplastic film is placed on top of the uncovered tissue section on a glass slide. A near-infrared laser pulse is transmitted through the plastic cap, causing the film to melt into the tissue. The cap is then lifted off of the tissue section, taking the captured cells with it. The cap is then inserted into an eppendorf tube containing appropriate buffer for molecular analysis by PCR. An image archiving workstation is attached to the system, allowing for complete documentation of the experiment.
- A Dell 400 workstation for image analysis and processing. This system contains software for image analysis and processing, as well as for final preparation of image figures for publication. Attached to the computer is an AGFA StudioStar flat-bed scanner and a Fujix Pictrography 3000 digital image printer. This Fujix printer produces photographic-quality digital prints, and is used for both publication images and poster images.
All of these instruments are linked via ethernet connections, allowing sharing of images within the facility, as well as transmittal of the digital images off-site. Extensive image processing and analysis software packages located on the various computers housed within the Cell Imaging Facility, including the central imaging workstation, can then be utilized for analyzing any image digitally captured. Moreover, images can be imported into desk-top publishing packages and prepared for publication printing on the Fujix video printer, or for slide transparency reproduction.
