The Microscopy Core of the Program in Membrane Biology (PMB) at MGH is equipped and staffed to provide a wide range of services to investigators from MGH and the Boston area scientific community in the area of light and electron microscopy. The Core is housed on the 8th floor of the Simches Research Center at MGH, and is directed by Dr. Dennis Brown, Ph. D. The Core will provide services ranging from complete performance of the technical procedures required, to full training of personnel. Among the techniques available are laser scanning confocal microscopy, spinning disk confocal microscopy, tissue fixation, sectioning, immunostaining and conventional immunofluorescence microscopy, image analysis, and all aspects of electron microscopy including immunogold staining. The Core facility provides a unique and important resource for members of the scientific community as they approach a variety of research problems that call for these advanced microscopy techniques.

Director: Dennis Brown, Ph. D.
Phone: (617) 726-5665
Fax: (617) 643-3182
Email: dbrown9@partners.org

Other Staff

Senior Lab Technologist: Robert Tyszkowski (Confocal Microscopy)
Phone: (617) 724-9694
Fax: (617) 643-3182
Email: rtyszkowski@partners.org

Senior Lab Technologist: Mary McKee (EM, Sectioning)
Phone: (617) 726-3696
Fax: (617) 643-3182
Email: mckee@helix.mgh.harvard.edu

Location of Core:

The facility is located at the Program in Membrane Biology, 8th Floor, MGH Simches Research Center, 185 Cambridge Street, Boston.

Major Equipment:

• JEOL 1011 electron microscope with fully digital image capture and processing
• BioRad/Zeiss Radiance 2000 confocal microscope
• Perkin Elmer LCI Spinning Disk Confocal Microscope
• Nikon Total Internal Reflection Fluorescence (TIRF) microscope system
• Nikon E800 Eclipse upright epifluorescence microscope with Hamamatsu Orca CCD camera and complete image analysis software (IP Lab)
• Nikon 80i upright epifluorescence microscope with Hamamatsu Orca CCD camera and complete image analysis software (IP Lab)
• Nikon TE300 inverted microscope with wide-field optics and Hamamatsu Orca CCD camera for fluorescence and ratio imaging, with Improvision OpenLab software
• Olympus BH2 upright microscope with Spot Insight color CCD camera and Macintosh G4 computer for light microscopy
• Olympus doubleheaded light microscope for section examination
• Reichert ultramicrotomes for routine EM sections (2)
• Leica FCS ultracryomicrotome for ultrathin frozen sectioning
• Microm cryostats (2)
• Leica microtome for paraffin sectioning
• MMI CellCut laser microdissection system
• Cressington freeze-fracture device (at CNY8 facility)
• Leica APS freeze-substitution system
• Several Macintosh G4-based computer workstations equipped with IP Lab Spectrum software for image analysis
• 2 Power Mac G5 based workstations with Improvision Volocity visualization, classification (morphometric analysis) and restoration (digital deconvolution) software, 8 gigabytes of RAM, and 256 mb Radeon video cards

1. Technical Services
The core will perform conventional immunofluorescence microscopy, confocal microscopy, live-cell imaging, conventional electron microscopy and EM immunogold labeling, tissue sectioning, tissue fixation, and computer based image analysis and quantification. In some cases, investigators will provide their own stained tissues/cell for imaging, while in others the Core will perform all services from fixation and sectioning, to staining and imaging.

2. Training Services
The core technicians will also provide training to high frequency users who wish to invest the substantial time necessary for learning to use the equipment proficiently.

3. Consultation Services
Dr. Dennis Brown and the Core technologists will be available for consultation in the use of the different microscopy techniques, and to provide technical advice, such as the different fixation techniques that enable antibodies to detect their antigens.

4. Development for Core Enhancement
Dr. Dennis Brown and the advisory committee will continually explore new techniques to be offered as part of the core service.

* Without a technician is an option available for frequent users who have undergone extensive training by the technician and for use of the confocal system during “off” hours (when technician is not available). It is also applicable to users of the image analysis systems for which technical help may not be required. Prices w/o technician are $30/h (MGH), $45/h (Non-MGH) and $80/h (Industry). The w/o technician option is not available for some services (e. g., EM, freeze-fracture and ultrathin frozen sectioning).

To access the core, please contact the appropriate persons above (Dennis Brown for general inquiries, Robert Tyszkowski for light and confocal microscopy, and Mary McKee for EM work and sectioning).

membranebiology.mgh.harvard.edu/