An open access facility, the Analytical Imaging Shared Resource is located in the Diabetes Research Institute and contains 1,031 square feet of laboratory, office, and storage space. The shared resource is equipped with state-of-the-art technologies and instrumentation including:
Leica MP-NDD4/SP5/FCS/FLIM multiphoton/confocal upright F-Techniques microscope (MP/SP5)
Includes Coherent Chameleon Ultra II Ti:Sapphire laser with 3.8 Watts peak average power, 4 external non-descanned detectors (NDD4), SP5 spectral confocal scanhead with high resolution and high speed resonant scanners, 405, 458, 476, 488, 496, 514, 561, 594, and 633 nm laser lines, ISS fluorescence correlation spectroscopy (FCS) with two avalanche photodiode detectors, Becker & Hickl time correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM), and three high performance internal photomulitplier tubes (PMTs). The resonant scanner enables 14 frames per second at 512×512 pixels for 2 channels, up to 210 frames per second for 512×16 pixels for 2 channels, or 8000 lines per second. The MP/SP5 is equipped with a full set of water saline immersion dipping lenses. Z-series, time lapse, and spectral series are available. This multiphoton microscope was one of the first in the world with Leica’s NDD4 non-descanned detector module for deep tissue imaging by multiphoton excitation fluorescence microscopy. The NDD4 module features simultaneous blue, cyan green, and red fluorescence acquisition. The Coherent Chameleon Ultra II Ti: Sapphire multiphoton laser has 3.8 Watts peak average power, enabling deep tissue imaging.
Leica SP5 spectral confocal inverted microscope (SP5 inverted)
Includes motorized stage, standard and high resolution Z-focus, SP5 spectral confocal scanhead with high resolution and high speed resonant scanners, 405, 458, 476, 488, 496, 514, 561, 594, and 633 nm laser lines. The resonant scanner enables 14 frames per second at 512×512 pixels for 2 channels, up to 210 frames per second for 512×16 pixels for 2 channels, or 8000 lines per second. The SP5 is equipped with a full set of oil immersion lenses. Z-series, timelapse, and spectral series are available. The high speed Z-galvonometer enables high speed XZ scans (200 um range), standard Leica DMI6000 Z-drive for large distance Z-series (limited by the opacity of your specimen), automated high resolution scanning stage with stage tiling and mark & find.
Leica AS LMD laser microdissection
Laser capture microdissection using the Leica AS LMD allows investigators to obtain essentially pure samples of a desired cell population from tissue sections for phenotypic analysis of dissected cells in their native state. The dissected cells can then be used to extract RNA, DNA, or proteins for further studies. Thus, LCM offers unprecedented access to specific cells, including cancer cells, for defining their pattern of gene expression, in combination with powerful techniques such as gene array and real-time PCR. The microscope has a motorized stage and camera that enables “specimen overview” images of entire tissue sections.
Leica DMIRB Inverted Microscope
Standard widefield fluorescence and brightfield “pathology” microscopy can be performed using the Leica DMIRB inverted microscope equipped to perform triple fluorescence, phase contrast and histology/immunohistochemistry light microscopy. We have upgraded our Leica DMIRB inverted microscope to a new Leica DFC495 color digital camera, Leica LAS image acquisition software, HP30w 2560×1600 pixel computer monitor and new Dell PC with fast RevoDrive solid state drive (SSD) PCI-Express card. The new DFC495 camera has much better color rendering than the old MicroPublisher RTV5 camera. The LAS software has an excellent stitching command.
MetaMorph Imaging System
The MetaMorph Imaging System (MIS) consists of hardware and software that enables the capture and analysis of microscopy or digital images obtained through the use of the instruments described above. MetaMorph offers more sophisticated capturing and analytical capabilities than the software provided with those instruments. The MIS can be used for live cell imaging, multi-label fluorescence, confocal microscopy, motion analysis, co-localization studies, FISH, FRET, FRAP, and live/dead cell assays. Most image analysis is done with MetaMorph, and the core can help users on projects involving Adobe Photoshop, NIH ImageJ, and additional software.
Pathscan4 microscope slide scanner
The Pathscan4 is the latest generation microscope slide scanner from Meyer Instruments. The Pathscan4 scans at 7,200 dpi (3.6 um pixel size) and can scan an entire brightfield histology or immunohistochemistry microscope slide in 1 minute. Users have the option of using the Quikscan software or LaserSoft SilverFast Ai.