Instrumentation:

Upgraded 5-Color FACScan Analyzers (2 available):

• A bench top analyzer with two lasers for fluorochrome excitation: The primary laser is a 15 mw argon (488 nm) laser and the secondary laser is a 25 mW red diode (637 nm) laser.
• Seven detectors: Two for light scatter (forward and 90°) and five for fluorescence. The optical filters for the fluorescence detectors are fixed at FL1 = 530±15 nm, FL2 = 585±21 nm, FL3 = 668 nm long pass, FL4 = 666±13 nm and FL5 = 740 nm long pass.
• Suitable for immunofluorescence (up to 5 color) and DNA content/cell cycle analysis.
• Commonly used fluorochromes (please note that Texas Red can not be used  as a single fluorochrome on these instruments):

                        Download a "Flurochrome Guide" pdf          

FACSVantage SE High Speed Cell Sorter:

• A dual laser instrument (argon and helium-neon lasers) capable of generating three simultaneous laser beams for fluorochrome excitation (UV at 340-360nm, blue at 488nm and red at 633nm). Please note that Texas Red can not be used as a fluorochrome with these lasers!
• Ten detectors: Two for light scatter (forward and 90°) and eight for fluorescence.  The fluorescence detectors are arranged so that  four receive signals from  the primary 488 nm beam, two from the second 633 nm beam and two from the third 340-360 nm beam. Optical filters may be changed on each of these fluorescence detectors to capture a particular fluorescence emission wavelength.
• Cell sorting capabilities for two populations defined by any combination of light scatter and fluorescence characteristics. The maximum cell analysis rate for sorting is 8 x 10⁶ cells/hour. To view more information about cell sorting click here.
• A recirculation water bath is available to maintain the sample tube and the sorted cells at a defined temperature.
• An Automatic Cell Deposition Unit (ACDU) option allows the sorting of defined numbers of cells into the wells of a microtiter or other tissue culture plate.

Data Analysis:

Computers:

        Each cytometer is operated by a Macintosh G4 computer equipped with a Fujitsu 235 Mb magneto-optical disk drive and 100 Mb ZIP disk drive for data transfer and storage. Those bringing disks for data storage should be sure to format them for Macintosh use.  Data analysis service is available by core lab personnel or investigators may use one of our off-line workstations to perform their own data analysis.  Off-line workstations are located at: 

• FACScan Lab in room 603 Wintrobe (available on first-come first-served basis). 
• Department of Pathology in room 5B124 SOM.
• Huntsman Cancer Institute, Samlowski Lab area in room 5254 (ask for John McGregor for help locating the computer).
• Eccles Institute for Human Genetics (EIHG), Zimmerman lab in room 4250A.

Software:

        Software available includes Becton-Dickinson's CellQuest, the basic data acquisition and analysis software,  on all off-line workstations.  In addition, available on the workstation in 603 Wintrobe is a DNA/Cell Cycle modeling program ModFit,  a more sophisticated flow cytometry data analysis program WinList, and an antigen quantitation program  QuantiCalc all three from Verity Software House.

Support Services:

        Full service by lab personnel is available for staining, analyzing and data analysis during normal working hours. Training is provided for individuals who wish to operate the FACScans themselves and take advantage of off-hour access. Collaborative support is also available for developing new assays and applications of the technology.