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The Microscope Imaging Facility (MIF) is located in the Health Sciences Center at West Virginia University. This facility provides access to advanced biomedical imaging technology including state-of-the-art light microscopes, as well as image analysis software packages. The Imaging Core Facility provides image acquisition and analysis services on a fee-per-use basis to the West Virginia University research and academic community. The facility provides individualized training for each user on the appropriate operation of the microscopes as well as downstream image analysis. The facility staff is available to help plan imaging experiments, ranging from advice on fluorophore selection to the development of new imaging routines such as FRAP or FRET for live cell experiments. The staff will collaborate more extensively with labs that have projects that are heavily dependent on microscopy. The facility will assist researchers in preparing data and methodology for publications and grant applications. Resources in this facility include:
MIF Nikon Live Cell: This system supports fluorescent and DIC imaging in live cells. This is a Nikon TE2000S inverted epifluorescent microscope with Prior filter wheels and a high sensitivity, high resolution Photometrics Coolsnap HQ CCD camera. It is controlled by the MetaMorph and MetaFluor software packages, which support high end image analysis. This system can do multi-color time-lapse, CFP/YFP FRET and calcium ratio imaging. There is a Bioptechs dish heater and objective heater to maintain a constant temperature in the sample. A tissue culture incubator is available to support the cells until the acquisition begins.
MIF Nikon Swept Field: This is a new Nikon Swept Field confocal system with 3 solid-state lasers (491, 561 and 638 nm) that is designed for high speed confocal acquisition in live cells. This system uses a Photometrics QuantEM CCD camera with on-chip gain to maximize signal and to increase the rate of acquisition. This system is also equipped with a laser TIRF (total internal reflection fluorescence) illuminator which is able to achieve very high sensitivity and resolution with fluorescent molecules in the plasma membrane at the interface with the glass dish. This system is equipped with an epifluorescent illuminator and has filter wheels and shutters to support multi-color time lapse acquisition. The Perfect Focus system maintains the focal plane during
movies. Both the TIRF and epifluorescent images are captured with high resolution Photometrics Cool-Snap CCD camera. This microscope setup is controlled by the Nikon Elements software package which also supports high end image analysis. There is a Bioptechs dish heater, objective heater and perfusion chamber to maintain a constant temperature in the samples. A tissue culture incubator is available.
MIF Olympus Histology: This system is an upright Olympus AZ70 epifluorescent/transmitted light microscope equipped with the MicroBrightField Neurolucida and Stereo Investigator software packages. This system is used for color histology records, 3-D neuron reconstruction, serial section reconstruction, anatomical mapping and morphometry analysis. It also has filter sets to capture blue, green and red fluorescent images.
MIF Zeiss Confocal: Confocal microscopy is a technique that removes out of focus light from images to improve the resolution and clarity of images. This system is a Zeiss LSM 510 laser scanning confocal on a Zeiss Axio Observer inverted microscope. It has 3 lasers with 4 laser lines (488, 514, 543 and 633 nm). There are 3 PMTs for collecting fluorescent light, as well as a separate PMT for collecting a DIC image of the cells. This system uses the Zeiss LSM software which supports z-slices and 3-D rendering and colocalization analysis.
MIF Zeiss PALM: This is a Zeiss PALM MicroBeam system for non-contact microdissection of single cells or groups of cells. It uses a UV laser to cut around the sample of interest, and then a defocused burst of laser energy is used to catapult the sample into the cap of a microfuge tube. The isolated cells can then be used for DNA, RNA or proteomic analysis. This system also has the capacity to collect cells under sterile conditions for further cultivation.
MIF Zeiss Violet Confocal: This system is a new Zeiss LSM 510 laser scanning confocal on an upright Zeiss AxioImager microscope. It has 4 lasers with multiple laser lines (405, 458, 477, 488, 514, 543 and 633 nm). The violet laser on this system allows users to image additional fluorophores, including DAPI and quantum dots. There are 3 PMTs for collecting fluorescent light, as well as a separate PMT for collecting a DIC image of the cells. This system uses the Zeiss LSM software which supports z-slices and 3-D rendering, time lapse acquisition, colocalization analysis and fluorescence recovery after photobleaching (FRAP) experiments.
For more information on the Microscope Imaginf Facility: http://anatomy.hsc.wvu.edu/mif/
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