Imaging Core

 

The HIHG Imaging Core offers UM colleagues and the South Florida scientific community a resource for microscopic imaging that is constantly updated and improved to remain cutting edge.

Currently installed:

  • Zeiss LSM 710 Quasar spectral confocal microscope.

Equipped with four lasers:

1.   405nm diode laser: for the ultraviolet-violet excitation spectral range dyes (DAPI, Hoechst, BFP);
2. 458nm/477nm/488nm/514nm Argon lasers: for blue excitation range spectral dyes (CFP/mTFP/GFP/YFP, Alexa Fluor 488);
3. 561 diode laser: for the green excitation spectral range dyes (DsRed; mCherry, mKate, Alexa Fluor 555 or 568)
4. 633 nm HeNe laser: for red excitation spectral range dyes (Alexa Fluor 647, Cy5).

The new Quasar spectral detector permits recording of X(YZT) lambda image stacks for the localized analysis of spectral signatures of the specimen and enables up to 10 emission signals to be precisely separated via software, even if the fluorescence spectra are overlapping. The ZEN 2009 software includes a spectral "Smart Setup" to help setup optimized image acquisition. Smart Setup uses the open data access PubSpectra dataset developed by Dr. McNamara, http://sylvester.org/AICF/PubSpectra.

The LSM710 microscope includes motorized XY stage and Z focus, with both stage tiling and mark & find capability for Z-series and/or time-series. 37 Celsius warming units are available for both 35 mm imaging dishes (e.g. from www.glassbottomdishes.com, FluoroDishes from www.wpi-inc.com,  Willco Wells from http://www.tedpella.com, CellView from www.greinerbioone.com) and for 24-well multiwell plates (e.g. Corning 3526 plastic bottom for low resolution imaging, P24G-1.5-13-F from www.glassbottomdishes.com for high resolution imaging).

If your specimen is to be imaged at room temperature, you can use a microscope slide or a 35 mm imaging dish, 50 mm imaging dish, or any SBS multiwell plate format (6, 12, 24, 48, 96, 384). Best results are obtained from image-quality dishes or plates cited above. For multiwell plates, consider Whatman ClearView or glass bottom plates (http://www.whatman.com/SpecialtyMicroplates.aspx, BD Falcon glass bottom plates http://www.bd.com/resource.aspx?IDX=1108, or Aurora Biotechnologies COP high quality plastic plates http://www.aurorabiotechnologies.com.

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Applications: high resolution cell and tissue imaging, Z-series, XY tile scanning, FRET, FRAP, time lapse live cell imaging with 37 C control, optional humidity, CO2 control.

Additional Microscopes:

  • Nikon bright field Eclipse 50i microscope, equipped with Qimaging MicroPublisher 5.0   RTV digital digital camera.
  • Nikon Eclipse TE2000-U fluorescent microscope equipped with Qimaging Retiga EXi  Fast 1394 digital camera.
  • Leica DMIL inverted microscope for cell culture equipped with Qimaging QICAM Fast 1394 digital camera. Dr. McNamara can arrange access to a multiphoton, fluorescence lifetime imaging, fluorescence correlation spectroscopy microscope (MP/FLIM/FCS) for anyone needing these capabilities.

For more information, please contact George McNamara, Ph.D. gmcnamara@med.miami.edu
Image Core Manager: Analytical Imaging Core Facility (AICF), DRI 6025
John P. Hussman Institute for Human Genomics, Diabetes Research Institute,
Interdisciplinary Stem Cell Institute, and UM/Sylvester Comprehensive Cancer Center