Flow Cytometry and Fluorescence Imaging Shared Resource

Sample Images

Click on thumnails for larger view

Director, Alice L. Givan, Ph.D.
Associate Professor of Physiology
Dartmouth Medical School

Contact Information

Location: Third Floor, Borwell West
E-Mail: givan@dartmouth.edu
Mail: Campus: HB 7700
Website: http://www.dartmouth.edu/~celllab

Co-Directors

Paul M. Guyre, Ph.D.
Charles P. Daghlian, Ph.D.

Staff

Kenneth A. Orndorff, M.S., Imaging Supervisor
Gary A. Ward, B.S., Flow Supervisor

Mission

The Cell Analysis Shared Resource provides an advanced instrumentation facility (the Herbert C. Englert Cell Analysis Laboratory) for the study of cells. It aims to:

Provide state-of-the-art flow cytometers and image-analysis systems.
Provide computer hardware and software for the analysis of cell data derived from these instruments.
Train scientists in use of the instruments and software.
Develop and implement new techniques for cell analysis.
Encourage communication among investigators about methods of common interest.

Facilities

The Herbert C. Englert Cell Analysis Laboratory occupies a suite of rooms on the third floor of the Borwell Research Building at the Dartmouth-Hitchcock Medical Center. The rooms house the flow cytometers and imaging systems; computers and printers for data analysis; desk space for staff; incubators; a tissue culture hood; a fume hood; and wet lab bench space for basic cell preparation. The Englert Laboratory also provides a flow cytometer for the Hanover campus; it is in the Molecular Biology Shared Resource on the second floor of the Vail Building. Currently the facility is used by more than 300 people, representing the labs of approximately 75 principal investigators, almost all of whom are members of the Cancer Center.

Equipment

For flow cytometry, the Laboratory is equipped with six Becton Dickinson instruments:
• A FACStar Plus cytometer with two lasers and a FACSAria cytometer with three lasers; both run by a dedicated operator for up-to fifteen parameter high-speed sorting and technically advanced methodologies.
• Two FACScan five-parameter cytometers (one in the Borwell Building and one on the Hanover Campus) run by individual users for routine analysis of cell surface, cytoplasmic, and nuclear antigens and for DNA analysis of ploidy and cell cycle stage.
• A FACSCalibur cytometer (with 488 and 635nm excitation) run by individual users for simultaneous analysis of two scatter and four fluorescence parameters.
• A FACSCanto cytometer, with two lasers and the ability to detect six fluorescence parameters.

The Laboratory also has three independent microscope systems:
• A Zeiss LSM 510 Meta Laser Scanning Confocal Microscope System with four lasers has the ability to obtain multi-color, 3-dimensional images of cells and tissues and to capture spectral information from the emitting fluorochromes.
• A Zeiss stereo/combi wide-field microscope that uses low magnification or a 10x or 20x objective to capture black and white or color digital images.
• An inverted wide-field microscope with the ability to look at fluorescence from cells, tissues, or the wells in a 96-well plate.

To bridge flow and imaging technologies, the Englert Laboratory has:
• A CompuCyte laser scanning cytometer (LSC) which scans cells or tissues on a slide and provides histograms and correlated two-color dot plots of fluroescence intensity distributions from the cells in the scanned area; these fluorescence intensity characteristics can then be mapped to locations on the slide in order to capture microscope images of individual cells with particular fluorescence properties.

There are, within the Laboratory, networked Macintosh and PC computer workstations with software for the analysis of flow and image data.

Services

The Cell Analysis Laboratory is supervised during normal working hours. Instruments are available 24 hours a day, 7 days a week for users who have been trained to work without supervision. A sign-up system operates for all instruments. Feasibility studies, training sessions, development of new methodologies, and data analysis on the computers are provided without charge.