FLOW CYTOMETRY FACILITY INSTRUMENTATION

ANALYTIC FLOW CYTOMETERS

Currently we provide five flow cytometers for analysis only work.

BD FACSCalibur has been modified by Cytek Development to provide 488nm and 635nm excitation and up to 5 colors of fluorescence. The FACS Calibur is in a BSL-2 room allows for the analysis of samples that contain or may contain viable infectious materials - this includes all human samples. BSL2 training and Schedule F required for operation.

Three Dako CyAns are available that each have 3 lasers for excitation (405nm, 488nm, and 635nm) providing up to 9 colors of fluorescence.

BD LSRII provides 7 lasers (355, 405, 488, 532, 561, 592, and 640nm) and 18 fluorescence detectors plus forward and side scatter. Forward scatter may be detected with either a photodiode or PMT.

NEW: Stratedigm S1000Ex cytometer for Microparticle Analysis. 3 Laser/13 color+ SSC and FSC size: 21"x21"x24" footprint Red Laser (640nm, 100mW, Solid--‐State) Blue Laser (488nm, 50mW, Solid--‐ state) Violet Laser (405nm, 100mW) No laser alignment required Particle detection from .2μ to 40μ Dedicated automatic clean and shut down procedures for the instrument Analysis rate: 10,000 events/sec Carryover:<0.1%, automatic backflush between samples Ability to measure true width/time of flight based on any trigger parameter.

All of these instruments may be investigator/staff operated after attending our training course or you may request that Facility staff operate the instruments for you.

The LSRII, and all the CyAns are not available for biohazardous samples. Any samples that may contain infectious agents must not be run on these instruments unless the samples are fixed with formaldehyde.

SORTING FLOW CYTOMETERS

Three sorting instruments are available.

We have our original MoFlo (purchased in 1997) still operating, the newer generation MoFlo XDP (installed in April 2007), and our most recent acquisition, the iCyt Reflection (installed November 2008). Each instrument provides unique configurations and strengths. The MoFlo provides for up to 3 laser excitations and 8 colors of fluorescence. The MoFlo has a tunable argon and a tunable krypton laser to provide a variety of laser lines along with a 633 He-Ne laser. The MoFlo XDP uses fixed wavelength lasers and can provide 3 excitations (405nm, 488nm, and 633nm) and 10 fluorescence detectors.

The iCyt Reflection provides 2 sorting units (HAPS) that can sort simultaneously thereby doubling throughput. The Reflection will direct up to 6 laser excitation beams (355nm, 408nm, 488nm, 532nm, 594nm, 640nm) into each HAPS. Eleven fluorescence detectors are available on each HAPS but higher numbers may be possible with PMT sharing. The MoFlos are available for cells not containing infectious agents. The Reflection is in a BSL-2+ room (operated using BSL-3 procedures) to provide sorting for all BSL level samples at or below BSL-2. The sorters are only available operated by Facility staff.

BD FACS Aria II SORP - 4-laser sorter for BSL-2 samples operated as the Reflection does. Includes cell indexing and FS PMT for microparticle sorting. Configuration link.

OTHER INSTRUMENTATION LOCALLY:

Amnis Image Stream - The Image Stream System collects images of cells that have been hydrodynamically focused in a flow cuvete with sheath as the background.  It collects high resolution digital images of cells in flow.  The Image Stream is located at Duke University at Duke University Medical Center in the Levine Sciences Research Building (LRC). Interested investigators should contact Gerry Blobe (gerard.blobe@duke.edu) or Mythreye Kathikeyan (mythreye.k@duke.edu). User fees apply. Click here for more information.