IMAGING (CONFOCAL, VISUAL/FLUOR SCOPES, IVIS)
The Imaging Core at Roger Williams Medical Center is comprised of the following instrumentation: a Zeiss LSM 700 confocal laser scanning microscope, two Zeiss Axioplan 2 fluorescence microscopes, an inverted Zeiss Axiovert 200M microscope, and a PerkinElmer IVIS Imaging System.
Zeiss LSM 700 confocal microscope:
The Zeiss LSM 700 microscope is a four laser, point scanning confocal with a single pinhole. It is in an inverted configuration to facilitate live-cell imaging. A key innovation of the LSM 700 system is the use of a variable secondary dichroic (VSD) beamsplitter, which enables efficient yet rapid detection of up to four different color signals. Rather than requiring separate filters for each desired fluorophore, this system uses a sliding dichroic mirror to rapidly direct specific wavelengths to selected photodetectors. This technology allows the use of the entire emission spectrum for imaging, while simultaneously preventing crosstalk between closely positioned fluorophore emission spectra.
LSM 700 Capabilities:
- Excitation lasers at 405nm, 488nm, 555nm, and 639nm
- 2 channel spectral detection
- Objectives: 10X/0.25, 20X/0.8, 40X/0.6, 63X/1.4 Oil DIC
Zeiss Axioplan 2 optical microscope:
The Zeiss Axioplan 2 imaging microscope is an upright modular transmitted light and incident light polarizing and fluorescence microscope with a pyramid design and an Infinity Color-corrected Optical System. It has available 5 objectives: 10x, 20x, 40x, 63X and 100x with capability for phase contrast, differential interference contrast (40x, 63x and 100x objectives) and fluorescence microscopy. Filter cubes are loaded for FITC/GFP, TRITC/Cy3 and dual-channel fluorescence. The stage is motorized in Z and the new Axiovision Release 4.7 software can control the stage, filter sets, and AxioCam camera to provide Z-stack and multi-channel digital image capture capabilities. These microscopes are capable of capturing monochrome images (standard fluorescent imaging mode, for taking pseudo-color single and multi-channel fluorescence images, or basic B&W bright field), and color images (for taking true-color images of H&E stains, etc.).
Zeiss Axiovert 200M optical microscope:
The Zeiss Axiovert 200M microscope is an inverted fluorescence microscope with motorized Z-axis acquisition and a filter wheel with Rhodamine, FITC, DAPI, and bright field. Objective lenses include 10x/0.3, 20x/0.4, 40x/1.3 oil, and 63x/1.4 oil DIC. The system is equipped with the AxioVision v4.6.3 acquisition software.
IVIS Model 100 Imaging System
The IVIS model 100 Imaging System is equipped with a Fluorescence Kit, an Anesthesia System, and the Living Image Acquisition/Analysis software. The primary use of this instrument is to non-invasively monitor, in real-time, bioluminescence and fluorescence within small animal models (e.g., rats and mice) and in vitro specimens (e.g., microtiter plates and culture dishes). The instrument can non-invasively monitor up to 5 mice simultaneously, and monitor in vivo multiple fluorescents and bioluminescent reporters simultaneously. The IVIS model 100 system is controlled by the Living Image Acquisition/Analysis Software Package, allowing for advanced quantification and analysis of all data acquired from each animal.
Applications of the system include:
- Tracking of bio-luminescent or fluorescently tagged cancer cells to monitor growth and metastasis.
- Monitor all forms of cell transplant experiments to quantify engraftment, cell growth, or cell differentiation, including studies of stem cell transplants, reconstitution of hematological and immune systems after irradiation.
- Monitor the in vivo gene activity in transgenic animals carrying appropriately tagged promoter indicator transgenes.
- Monitor in vivo gene activities in cells that have been reconstituted to carry a bioluminescent or fluorescently tagged expressed gene.
- Monitor any form of biochemical activity that can be assayed using a bio-luminescent or fluorescently tagged substrate.