Equipment

The SDU Flow Cytometry Core Facility is equipped with two analytical flow cytometers and a high-speed cell sorter as well as equipment for magnet-based cell separation.

The analysers:

Becton Dickinson FACScan Flow Cytometer
A basic 5-parameter cytometer for data acquisition, with or without assistance from core personnel. (Inexperienced users will be required to take a short training course prior to independent operation.)
    The FACScan has a fixed 488nm Argon laser and three-colour detection capabilities covering a substantial range of fluorochromes (including fluorescein isothiocyanate (FITC), green fluorescent protein (GFP), carboxyfluorescein succinimidyl ester (CFSE), 2,7-dichlorofluorescein diacetate (DCFD) or dihydro-rhodamine (DHR) as the first fluorochrome (FL1), R-phycoerythrin (PE) or propidium iodide (PI), as FL2, and Ethidium Bromide (EB) Peridinin chlorophyll protein (PerCP) or the PE-Cyanin 5 tandem (PE-Cy5) as FL3.

Becton Dickinson FACSCalibur Flow Cytometer
A dual-wavelength analytical flow cytometer equipped with an argon laser, for excitation at 488 nm, and a red diode emitting at 635nm. The FACSCalibur, is capable of four-colour analysis covering all the fluorochromes detectable with the FACSCan and, in addition, allophycocyanin (APC), Cy5 or Alexa Fluor 633 (as FL4).
     Both FACScan and FACSCalibur operate with closed fluidic systems, so use with biohazardous samples is possible with appropriate precautions.

The cell sorter:

Becton Dickinson FACSVantage/Diva Sorting Flow Cytometer
A FACSVantage SE cell sorter with digital electronics (FACSDiva option), equipped with two lasers; an Enterprise IIC argon laser providing 488 and 351-364 nm excitation and a Spectra Physics He/Ne laser emitting at 647 nm. The instrument has eight detectors (2 light-scatter and 6 fluorescent). The sorter operates with all the above-mentioned dyes, while UV excitation enables additional detection of the DNA dyes, Hoechst 33342/33258 and DAPI, or the Calcium-ion probe Indo-1. The cytometer is equipped for two-way and four-way sorting, as well as for single cell cloning. Sorting rates of up to 30,000 cells per second are achievable.

 Software and Data Processing:

The FACSCan and FACSCalibur flow cytometer are supported by Cell Quest software running on MAC computers. The FACSVantage/Diva cell-sorter employs PC-based Cell Quest Pro software. Both PC and MAC workstations are available for data processing.

Magnet-based cell separation:

The core facility is also equipped for magnetic sorting using the Miltenyi VarioMACS and SuperMACS systems. These enable rapid enrichment of cell sub-populations on a large scale (i.e. from samples containing up to 1011 total cells), which can be of great benefit as a preliminary step in the fluorescence-based sorting of very rare cell-types. Prices for the use of this technique are available on application to Graham Leslie (gleslie@health.sdu.dk).