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Flow cytometry

Flow cytometry is an essential technique for many cell biology or immunology applications. The KIR flow cytometry facility is managed by Jonathan Webber under academic supervision of Prof Lynn Dustin, and supports around 70 active users.

Flow cytometry manager

Equipment

The KIR flow facility operates 2 BD analysers (LSRII, Fortessa X20), 1 Cytek spectral analyser (Aurora), and 1 BD cell sorter (Aria III).

The KIR flow facility also provides a Meso Scale Quickplex SQ120 imager, for detection of up to 10 soluble analytes simultaneously.

  • LSR II (R/B/V), Fortessa X20 (R/B/V/YG) are 14-colour maximum panel size. The YG laser on the Fortessa offers an advantage with dsRed/mCherry/tdTomato reporters. The Fortessa has a plate loader.
  • Aurora (R/B/V/YG/UV) is a spectral cytometer capable of a 40-colour panel size, with a very effective plate loader for automation. A major part of the KIR scientific remit is immune/stromal crosstalk. This is facilitated by autofluorescence extraction available on the Aurora, whilst the large panel size enables multiplexing of several smaller panels.
  • Aria III (R/B/V/YG), configuration equivalent to the Fortessa. Cell sorter is enclosed in a BSC for safe sorting of unscreened blood. Can sort 4 populations simultaneously or into single wells of a 96-well plate. Maximum throughput around 50 million cells/hr for PBMC/splenocyte or similar.

Support 

The flow facility provides Flowjo portal licenses for data analysis. The facility manager can assist with experiment design, troubleshooting and analysis. If unsure, please email before reagent purchase!

Cell sorting is operator-run in the first instance. The flow manager will discuss your experimental requirements (viability, downstream processing, input number and expected yields, duration, biosafety) prior to scheduling, in order to maximize likelihood of good experimental outcome.

Booking and access

Internal booking is available via BookIt once training has been completed. Please contact the flow manager to arrange training and access. External access (including sorting) is possible via contacting the flow manager, though internal usage is prioritised.