THE HERBERT C. ENGLERT CELL ANALYSIS LABORATORY
A SHARED RESOURCE OF THE NORRIS COTTON CANCER CENTER
DARTMOUTH MEDICAL SCHOOL
THIRD FLOOR BORWELL WEST, LEBANON, NH 03756
and ROOM 241, REMSEN, HANOVER, NH 03755
TEL 603-650-7661 or 7907
e-mail: givan@dartmouth.edu
The Englert Cell Analysis Laboratory is located in a suite of rooms on the third floor of the Borwell Building at Dartmouth-Hitchcock Medical Center. In addition, an "annex" is located on the second floor in the Remsen Building on the Hanover Campus.
For FLOW CYTOMETRY, the Lab is equipped with:
• a Becton Dickinson (San Jose, CA) FACStar Plus high speed sorting cytometer with two lasers (UV and 488 or 514 nm), run by a dedicated operator for six-parameter sorting (at rates up to 20,000 cells per second) and for technically advanced methodologies (for example, studies of pH change and calcium flux).
• two FACScan cytometers (one in Borwell with 5 parameters and the other in Remsen with seven parameters), and one FACSCalibur (6-parameter) cytometer, (all Becton Dickinson, San Jose, CA) run by individual users for routine analysis of cell surface, cytoplasmic, and nuclear antigens, for fluorescence from cell tracking dyes, and for DNA analysis of ploidy and cell cycle stage.
• NEW: a Becton Dickinson FACSAria sorting cytometer, with three lasers (407 nm, 488 nm, and 633 nm) and the ability to detect forward and side scatter plus thirteen (13) fluorescence colors; this instrument is run by a dedicated operator and can sort at rates up to 30,000 cells per second with sorting of up-to four sub-populations simultaneously.
• NEW: a FACSCanto benchtop cytometer capable of detecting 6 fluorescence colors (plus forward and side scatter).
The Laboratory also has IMAGE ANALYSIS systems:
• a Zeiss LSM 510 Meta laser scanning confocal system with four lasers for fluorescence excitation (405, 458, 477, 488, 514, 543, and 633 nm) and two conventional PMT detectors and one multi-spectral meta detector for simultaneous analysis of up to eight fluorescence colors from cells on slides, in wells, or on plates.
• NEW: a Zeiss stereo, wide-field, fluorescence microscope, capable of acquiring digital images at low (stereo) magnification or with a 10x or 20x objective.
To bridge flow and imaging technologies, the Laboratory has a LASER SCANNING CYTOMETER:
• the CompuCyte (Cambridge, MA) laser scanning cytometer (LSC) scans cells or tissues on a slide and provides histograms and correlated two-color dot plots of fluorescence 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.
For COMPUTATIONAL ANALYSIS, there is, within the Laboratory, a computing area with networked Macintosh and PC computers and a Silicon Graphics workstation. Image analysis software includes Volocity by Improvision (for rendering and seeding and for quantitative co-localization and volume analysis), AutoDeblur (for deconvolution), NIH-Image/ImageJ, and Photoshop. Software for flow cytometric analysis includes CellQuest, ModFit, WinList, FCSPress, and FlowJo.
For PRINTING and DATA BACKUP, there is a dye-sublimation printer for publication-quality color graphics and a CD-ROM burner for archiving of files.
The Laboratory also has WET LAB space, with centrifuges, incubators, and a tissue culture hood for cell culture and preparation.
The Englert Cell Analysis Laboratory was established with an equipment grant from the Fannie E. Rippel Foundation and has received subsequent grants from the Rippel Foundation, the NIH Shared Instrument Grant Program, and Dartmouth Medical School. It is maintained in part by user fees and in part by the Core Grant from the National Cancer Institute to the Norris Cotton Cancer Center.
Alice Givan, Director
Paul Guyre and Charles Daghlian, Co-Directors
Ken Orndorff, Imaging Supervisor
Gary Ward, Flow Supervisor