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.
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.
Useful links
We have curated a comprehensive list of useful links of all things flow cytometry.
Publications
-
Elevated type-17 cytokines are present in Axial Spondyloarthritis stool
Journal article
BOWNESS P. and BROUGH I., (2024), Discovery Immunology
-
A first in disease phase 2, randomised, controlled trial of Granulocyte-Macrophage Colony-Stimulating factor neutralisation for Axial Spondyloarthritis (NAMASTE study)
Journal article
BOWNESS P., (2024), Lancet Rheumatology
-
Publisher Correction: Semaphorin 3A causes immune suppression by inducing cytoskeletal paralysis in tumour-specific CD8+ T cells.
Journal article
Barnkob MB. et al, (2024), Nat Commun, 15
-
Obesity Differs from Diabetes Mellitus in Antibody and T Cell Responses Post COVID-19 Recovery.
Journal article
Ali M. et al, (2024), Clin Exp Immunol
Latest publications
-
Elevated type-17 cytokines are present in Axial Spondyloarthritis stool
Journal article
BOWNESS P. and BROUGH I., (2024), Discovery Immunology
-
A first in disease phase 2, randomised, controlled trial of Granulocyte-Macrophage Colony-Stimulating factor neutralisation for Axial Spondyloarthritis (NAMASTE study)
Journal article
BOWNESS P., (2024), Lancet Rheumatology
-
Publisher Correction: Semaphorin 3A causes immune suppression by inducing cytoskeletal paralysis in tumour-specific CD8+ T cells.
Journal article
Barnkob MB. et al, (2024), Nat Commun, 15
-
Obesity Differs from Diabetes Mellitus in Antibody and T Cell Responses Post COVID-19 Recovery.
Journal article
Ali M. et al, (2024), Clin Exp Immunol
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.