The Flow Cytometry Facility at the Dunn School

This facility is located in the Old School Buildingof the Dunn School in room 214.20.06. The facility comprises of an Analysis Lab and a Sorting Lab run by Nigel Rust (nigel.rust@path.ox.ac.uk) helped by Gabriela Ridlova

Flow Cytometry - What is it?

The Analysis Lab contains two Becton Dickinson and one Dako Flow Cytometer:

* A FACSort (4 colour)
* A FACSCalibur (4 colour)
* A Dako Cyan (9Colour)

The primary laser for all these instruments emits 488nm light, this will excite fluorochromes like FITC( fluorescein), PE (phycoerythrin) and PerCP (peridinin chlorophyll protein) or PE-Cy5 tandem conjugate (Tri-Color ,CyChrome). The 4 colour excited by diode HeNe laser (2nd Laser) is APC (allophycocyanin). For the Dako Cyan the 2^nd Laser allows the use of APC Cy7. The 3^rd Laser( on the Cyan) is 405 nm and can be used to excite Pacific blue and Pacific Yellow (or equilavents)

The FACSort uses CellQuest 3.3 software, the Calibur uses CellQuest Pro and the Cyan uses Summit 4.4 software on a PC.

The department holds a site licence for FlowJo, this will enable the analysis of data from all the instruments on your own PC/Mac. Please speak to Nigel prior to downloading this software>

The Sorter Lab contains two Dako Cytomation MoFlo Legacies,

These are modular instruments and have 3 lasers emitting at the wavelengths of 488 nm, 633 nm and 351-63 nm 9 or405nm) allowing the excitation of the common flow cytometry fluorochromes. This instruments are High-Speed cell sorters capable of throughputs in excess of 30,000 cells per second. The 4 Way sort module, enables us to sort 4 separate populations from one sample at once, the module CyClone provides for the isolation of single cells (or fixed numbers) and deposit them into various tissue culture plates (72 well Terasaki to 1536 well plates). The multi laser capability in combination with pulse processing and temperature control enables us to study the kinetics of cells in response to stimuli (like Ca2+ flux) in association with specific subpopulations markers.