Our Science – Garfield Website
Susan H. Garfield, M.S.
The facility provides Laser Scanning Confocal Microscopy using either a Zeiss LSM 510 UV system, a Zeiss LSM NLO (2-photon) Meta system or a Zeiss/Bio-Rad MRC 1024 system. Confocal software allows simultaneous or sequential three channel (24 bit) acquisition of 2D, 3D and 4D data. Three additional processing stations are available for post-acquisition analysis and publication quality prints can be produced using Codonics NP-1600 dye sublimation printers or Epson photo quality printers.
Imaging of both fixed specimens and live cells is provided. For live cell imaging, cells are placed in a temperature-controlled, perfusable enclosed chamber or an open chamber allowing acquisition over several minutes to several hours. Fluorescent Recovery after Photobleaching (FRAP), Fluorescence Resonance Energy Transfer (FRET), and co-localization studies can be conducted as well as imaging of UV excitable dyes. The IR laser of the 2-photon system facilitates imaging of thick specimens and minimizes tissue damage while the Meta detector provides multi-spectral imaging. Several software packages are available to users including Bitplane's Imaris for volume/surface rendering, tracking and measurements; Media Cybernetics' Image Pro Plus and 3D constructor; NIH's ImageJ and Adobe's Photoshop.
The core is supported by the Center for Cancer Research at no charge to individual users. Collaborations with outside laboratories are also considered, time permitting. Please refer to the publication list for examples of confocal imaging that have been done in this facility.
Investigators should contact Susan Garfield, Facility Manager, to discuss their project and facility procedures. If the project is appropriate, appointments will be made for using the confocal. The 'Confocal Slide Submission Form' found at http://elsie.nci.nih.gov/confocal is completed when samples are ready for imaging. Consultations on experimental design with emphasis on proper controls for co-localization, FRET and FRAP analysis is provided along with suggestions for optimum fluorophore selection, sample preparation and fixation, and sample mounting and storage until image acquisition.
To learn more about the services of the Core Facility please contact Susan Garfield.
This page was last updated on 6/10/2013.