Sylvester Comprehensive Cancer Center

Flow Cytometry Technologies

LSR-II

LSR-II, 8-color, flow cytometric analyzer- a sophisticated analytical instrument, used for complex analysis of cell samples. It has four lasers and is capable of 8-color analysis.

The Flow Cytometry Core Facility is housed in a 700-square-foot open-access/limited-access facility. The core is equipped with the following flow cytometers and instruments:

  • BD FACS. Canto-II, 8-color, flow cytometric analyzer
  • BD LSR-II, 10-color, flow cytometric analyzer
  • BD LSR-Fortessa-HTS, 19-color, high-throughput, flow cytometric analyzer
  • BD FACS Aria-I, 8-color, flow cytometric cell sorter
  • BD FACS Aria-IIu, 17-color, flow cytometric cell sorter, housed in a Baker BioProtect-II, BSL-2 bio-safety cabinet
  • BD FACS Aria-II, 10 color, flow cytometric cell sorter
  • MoFlo XDP, 7-color, high-speed cell sorter, housed in a Walker custom bio-safety cabinet
  • Zeiss fluorescence microscope
  • Invitrogen Countess cell counter
  • Partec CyFlow Space, 6-color flow cytometric analyser and absolute cell counter

Access to the Core’s instruments

Individual users can be trained independently to operate the Canto-II flow cytometric analyzer. With advanced training, select users also independently may operate the LSR-II and LSR-Fortessa-HTS analyzers. Cell sorting is performed with the two FACS Aria cell sorters and only by the facility’s operators. Sorting is available on-demand, for after-hours and weekend operation. The MoFlo XDP sorter is located in the new Stem Cell Institute, on the 9th floor of the Biomedical Research Building.

Instruments

Analysis

  • FACS Canto-II Analyzer – The three laser, fully digital FACS Canto-II analyzer is used for up to eight-color (ten-parameter) analysis of cell samples. It has a PC work-station, which runs the DiVa-6 software.
  • LSR-II Analyzer – The LSR-II analyzer is a sophisticated analytical instrument, used for complex analysis of cell samples. Recently fully upgraded, it now has four lasers and is capable of 10-color analysis. This fully digital instrument is equipped with a PC which runs the DiVa-6 software.
  • LSR-Fortessa-HTS analyzer – The new Fortessa analyzer also is a sophisticated analytical instrument, used for complex analysis of cell samples. It has five lasers and is capable of 19-color analysis. This fully digital instrument has a PC running the DiVa-6 software. It also is fitted with a HTS high-throughput automated sample loader, for rapid analysis of stained cells directly from culture plates.
  • Partec CyFlow Space analyser – The CyFlow Space is a compact, basic, pump-driven, two laser analyser, for six colour analysis and absolute cell counting. It has a PC which runs the FloMax software.

Sorting

  • FACS Aria Cell Sorters – The three FACS Aria cell sorters are used for complex analysis and sorting of cell samples. The instruments incorporate a fixed-alignment flow cell for enhanced sensitivity of fluorescent detection and ease of optical alignment. All Arias have “Whisper” aerosol evacuation systems fitted and are fully digital. They have PCs which run the DiVa-6 software and are interfaced to printers.
    • BD FACS Aria-I is a standard high-speed cell sorter. It has 3 lasers and is capable of up to 8-color simultaneous fluorescence analysis. No optical alignment is required and drop-delay calculation is automated. This is a good, basic-level sorter.
    • BD FACS SORP Aria-IIu is also a high-speed cell sorter, with an Automatic Cell Deposition Unit (ACDU) and temperature control for sorting. It has 5 lasers for up to 17-color analysis and cell sorting. This instrument is housed in a Baker BioProtect-II bio-safety cabinet, which enables sorting of BL-2 level samples. Again, no optical alignment is required and drop-delay calculation is automated. This is a custom-built (SORP), advanced-level sorter.
    • BD FACS SORP Aria-II high-speed cell sorter has 4 lasers and is capable of up to 10 color simultaneous analysis.
  • MoFlo XDP Cell Sorter – The BCI MoFlo XDP is an ultra high-speed (c.50,000/sec.) cell sorter, with three lasers seven fluorescence detectors, a CyClone plate deposition robot, temperature control and all housed within a custom-built Walker bio-safety cabinet. Daily optical alignment is required and drop-delay calculation currently is manual. This is an excellent, “old school” high-speed sorter.

Other

  • Zeiss Fluorescence Microscope – The Zeiss Fluorescence Microscope has a camera attachment and is provided for examination of fluorescent cell samples, before and/or after cell sorting.
  • *Invitrogen Countess cell counter – This uses just 10ul. of sample, to provide a total cell count, plus analysis of cell viability using Trypan Blue dye.

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