The Flow Cytometry Facility at the Dunn School

This facility is located in the Old School Building of 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)

Flow Cytometry - What is it?

Beginners Guide

The Analysis Lab contains one Becton Dickinson , Beckman Coulter , Cytek DxP8 Flow Cytometer’s and an Amnis ImageStream:

* A Cytek (8 colour 3 laser)
* A FACSCalibur (4 colour 2 Laser)
* A Dako Cyan (9 Colour 3 Laser)

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 Beckman Coulter Cyan and Cytek DxP the 2^nd Laser allows the use of APC Cy7. The 3^rd Laser ( on the Cyan and DxP) is 405 nm and can be used to excite Pacific blue and Pacific Orange (or equilavents)

The Calibur use’s CellQuest Pro , the Cyan uses Summit 4.4 software on a PC and the Cytek use’s and adaption of FlowJo.

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.

*Amnis ImageStream

This Instrument bridges the gap between Image Analysis and Flow Cytometry. By using a camera instead of photomultiplers it enables the image of a cell to be investigated with the advantage of Flow Cytometry to provide statistically enumeration. We have a 2 Camera system with 4 lasers to enable 12 channel data to be recorded.

Useful Information and forms

The Sorter Lab contains a Beckman Coulter MoFlo Legacy,

The Legacy is a modular instruments and have 3 lasers emitting at the wavelengths of 488 nm, 633 nm and 405nm) allowing the excitation of the common flow cytometry fluorochromes. This instruments is a High-Speed cell sorter 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.