Published on New York State Department of Health, Wadsworth Center (https://wadsworth.org)

Advanced Light Microscopy & Image Analysis Equipment

  • Confocal microscope Leica TCS SP5: Equipped with high efficiency photo-multipliers capable of six channel simultaneous detection (5 fluorescent and 1 bright field) and an acousto-optical beam splitter (AOBS). The ABOS has significantly higher transmission and is much faster than more traditional dichroic mirrors. This system collects more photons than any other technique - the result is bright, noise-free images and extremely low photodamage at high speed. Currently, we have separated 5 channels in a very challenging sample. A few of its features are highlighted below:
    • True confocal point-scanning - real optical sectioning
    • High dynamic sensors
    • Low dark current PMTs
    • Free tunable emission bands
    • Any dye adaptable
    • Spectral un-mixing- Dye separation: sufficient for 5 channels simultaneously
    • Spot-bleaching FRAP
    • 8 different laser lines available: 458, 476, 488, 496, 514, 543, 594, 633 nm
    • Objective lenses 10X to 63X
       
  • Auto-Slide Stainer, BioCare IntelliPATH: One of the greatest sources of variability in microscopy & pathology is sample preparation. Removing the human element as much as possible will reduce errors. Reducing variability and therefore increasing confidence in test results are critical to both public health and basic research. This system greatly reduces variability and improves efficiency through automation.
    This intelliPATH is a flexible immunohistochemistry (IHC) staining system with walk-away capabilities. Its unique technology enables small volumes of reagent (as little as 100 µL per slide) to be uniformly applied over the tissue sections on a slide. The intelliPATH offers continuous batch processing, allowing independent start and finish times for each batch of 10 slides as well as different protocols on each of 30 slides. This system can be used to automate slide, cell and section preparation, which can take days per run if prepared manually.
     
  • Wide Field Imaging Workstation:
    • Nikon TE2000
      • Fluorescence, Phase, DIC and Bright field imaging modes
      • 1, 2, 4, 10, 20, 40, 60, 100X Objective lens
      • Filter wheels for Dapi, FITC (GFP), TRITC (Texas Red), CY-5
      • Temperature controlled (RT → 50ºC) cell chamber for imaging over extended time periods
      • Cooled CCD camera (Roper HQ)
      • Z-stepper drive
      • FRAP (fluorescence recovery after photo bleaching) capabilities
      • Software for acquisition, post-processing, feature extraction, alignment, overlays, tracking and much more
    • Olympus X-70
      • Fluorescence, Phase, and Bright field imaging modes
      • 2, 4, 10, 20, 40, 60, 100X Objective lens
      • Filter cubes for Dapi, FITC (GFP), TRITC (Texas Red)
      • Temperature controlled (RT → 50ºC) cell chamber for imaging over extended time periods
      • Cooled CCD camera (Apogee Alta)
      • Z-stepper drive
      • Software for acquisition, post-processing, feature extraction, alignment, overlays, tracking and much more
         
  • Optical Projection Tomography (OPT) Workstation: There are many techniques for 3D imaging such as confocal microscopy. Including wide-field florescence microscopy, CAT scan, MRI, atomic force microscopy, OPT has the advantage of allowing large objects (1 cm3) to be imaged at higher resolution than CAT or MRI. OPT allows the specimen to be imaged both with transmitted as well as fluorescent illumination. With image algorithms, some borrowed from CAT and MRI imaging, this technology has a lot of room to evolve and improve both in terms of detectability and specimen type and size. Specimens (either mounted in glass tubes or native) are imaged in a rational manner and then projected into a 3D volume. See gallery for some really cool movies.
    • Fluorescence, Phase, and Bright field imaging modes
    • 2, 4, 10 Objective lens
    • Filter cubes for Dapi, FITC (GFP), TRITC (Texas Red)
       
  • Upright Fluorescent Scope: The software controlling the camera is easy to use, flexible and highly intuitive, and a DVD burner is available on this system. This system also has the capacity to perform calibrated measurements from simple distances to complex area/perimeter determinations.
    • Fluorescence, Phase, and Bright field imaging modes
    • 10, 20, 40, 60X Objective lens
    • Temperature controlled (RT → 50ºC) cell chamber for imaging over extended time periods
    • 3-shot Color Cooled CCD camera
  • Upright Polarized light Scope: Polarized light microscopy can produce both qualitative and quantitative measurement from birefringent specimens (decomposition of a ray of light into two rays when passing through certain samples). These specimens can be alive, fixed or inanimate. The colors in the image are a result of the interference between light waves as they pass through the specimen. The software controlling the camera is easy to use, flexible and highly intuitive, and a DVD burner is available on this system. This system also has the capacity to perform calibrated measurements from simple distances to complex area/perimeter determinations.
    • Polarized light, fluorescence, phase, and bright field imaging modes
    • 2.5, 5, 10, 20, 40, 60X Objective lens
    • 3-shot Color Cooled CCD camera
  • Stereo Fluorescent Scope:
    • Fluorescence and bright field imaging modes
    • 20-200X Zoom
    • Various illumination modes including transmitted and reflected
    • 3-shot Color Cooled CCD camera
       
  • Dual-head Stereo Scope: This is an ideal teaching microscope allowing two individuals to view the same field-of-view simultaneously.
    • bright field imaging modes
    • 20-100X Zoom
       
  • Microtome (vibrating-blade) Leica VT1000 Instrument:
    • Linear sectioning speed adjustment
    • Linear sectioning frequency adjustment
    • Sectioning thickness totalizer
    • 5 Different amplitude settings
    • Freely programmable sectioning window
    • Temperature from 4º - 27º C
    • Easy mounting and removing of knife holder and buffer tray.
    • Double-walled buffer tray providing excellent contrast
    • Cold light source and fiber-optical lighting system
       
  • Miscellaneous Equipment:
    • Cell viewing chambers and thermal regulation: Our laminar flow perfusion chambers allow cells cultured on coverslips to be followed at the highest possible resolution while maintaining the ability to rapidly change the bathing media at any desired time, i.e. for fixation or stimulation (Rieder and Cole, 1998). Our Rose chambers allow for long term (up to 3 day) LM studies. Cells can be maintained on any of the microscopes (listed above) with chamber heaters designed and built in house (Rieder and Cole, 1998), which can maintain temperatures between 0o-50oC for long (> 3 day) periods.
    • Laminar flow hoods and Incubators: For maintaining and growing various cell lines used in LM studies.