Flow Cytometry

Flow cytometry is a powerful tool for the multiparameter analysis of cells of all types. Three high powered high speed flow cytometers are available as well as one simpler cytometers and a three colour LED upright Fluorescence microscope in the Blizard Institute. The Blizard Institute instruments include a four laser 16 fluorochrome BDAria IIIu cell sorter and a five laser Cytek Aurora spectral analyser with 40 fluorochrome channels, five laser 18 fluorochrome analyser a BD LSRII, a three laser 8 fluorochrome BD FACSCanto II and a 3 laser 13 fluorochrome ACEA Biosciences Novocyte 3000.

Training and assistance will be provided by the Flow Cytometry Core Facility Manager.

NEW MIFlowCyte criteria for good practice in data publication, see Cytometry the scientific journal for the International Society for the Advancement of Cytometry, ISAC, see details and download form in Flow Cytometry Guides. All such data for publication in Cytometry has to be uploaded to FlowRepository.org

Sorter COSHH Form

All cell samples for sorting must be assessed for pathogenic and GM potential. Thus it is also required to supply a BioCOSHH form assessment of the cells due for sorting. Fixed samples can be sorted after the fixative is removed. More information can be found on the link, biological hazard. Cell sorting assessment form http://www.icms.qmul.ac.uk/flowcytometry/contacts/biologicalhazard/FACS%20Cell%20Sorting%20Assessment.pdf), Cell Sorting Biosafety Guidance, and QMUL Biological Safety Procedures documentation.

NEW ASSAY Design & Expert Assistance!

A REMINDER to all staff that the Flow Cytometry Core Facility offers NEW ASSAY DEVELOPMENT expertise, click here for list of new assay's already developed. Flow cytometry may be the answer to a research project problem and we can investigate whether flow cytometry can be used to investigate your research demands.

Expert support and technical assistance is also available to help with the design and development of experiments. Please see contacts for help and advice. Academic Lead for this core facility is Pro Dan Pennington, lead for the Centre for Immunobiology.

NEW BOOKING SYSTEM FOR 2017 Blizard Institute use only:- Please pre-register and add grant code to your account and click here for on-line booking with iLabs https://qmul.corefacilities.org/signed_out

Costs
Facility News
Contact Us
Instruments
Publications

ICMS Staff and associated Institutes

£30 per hour for BDLSR II, BDCanto II and ACEA Biosciences Novocyte 3000 analysis

£10 per hour for the Millipore Muse

£50 per hour for assistance

£50 per hour for cell sorting

External Academic

£50 per hour for analysis

£85 per hour for assistance

£100 per hour for sorting

Industrial

£70 per hour for analysis

£120 per hour for assistance

£175 per hour for sorting

Aria I upgrade in Feb 2014 to Aria IIIu - new Yellow-Green 561nm laser with 16 colour detection - purchased by User Funds
ACEA Bioscience Novocyte 3000 with Blue, Red and Violet lasers allows the collection of 13 fluorochromes, 5 from the Blue laser, FITC, PE, PE-CF595, PerCP or PerCP-Cy5.5, PE-Cy7, Red laser fluorochromes are APC & APC-Cy7, Violet laser fluorochromes, BV421, BV511, BV570, BV605, BV650 & BV780 - purchased User Funds 2015
SFIF funding will allow the purchase of a 5 laser, 40+ fluorochrome flow cytometric analyser - coming in Summer 2020

Dr Gary Warnes, Flow Cytometry Core Facility Manager
Tel: 020 7882 2402
email: g.warnes@qmul.ac.uk

Dr Gerard Hernandez, Deputy Flow Core Facility Manager
Tel: 020 7882 2402
g.hernandez@qmul.ac.uk

Prof Dan Pennington, Research Lead, Centre for Immunobiology
Tel: 020 7882 2302
email: d.pennington@qmul.ac.uk

Blizard Users for on-line booking please go to iLabs https://qmul.corefacilities.org/signed_out

BDFACS Aria IIIu - Four laser with 16 colour detection cell sorter, Blue Laser detects FITC or GFP or AF488, PerCP or PerCP-Cy5.5, Red laser detects APC, AF700, APC-Cy7, Yellow-Green laser detects PE, PE-CF595, PE-Cy5 or PE-Cy5.5, PE-Cy7, Violet laser detects BV421, BV511, BV650, BV650, BV711, BV780. The instrument will sort into 6, 12, 24, 48, 96 and 384 TC well plates with single cell index sorting available.
BDLSRII - Five laser 15 colour flow cytometric analyser with Blue laser which detects FITC & PerCP-Cy5.5 PE-CF595, Yellow-Green laser which detects PE, PE-Cy5, PE-Cy5.5 or PE-Cy5.5 & PE-Cy7, Red laser detects APC, AF700 & APC-Cy7, Violet laser detects BV421, BV511, BV605, BV650, BV711 & BV780 and UV laser detects DAPI and BUV737
BDFACS Canto II Three laser flow cytometric analyser with Blue laser which detects FITC, PE, PercP or PE-Cy5 & PE-Cy7, Red laser detects APC & APC-Cy7, Violet laser detects BV421 & BV511
ACEA Biosciences Novocyte 3000 with Blue, Red and Violet lasers allows the collection of 13 fluorochromes, 5 from the Blue laser, FITC, PE, PE-CF595, PerCP or PerCP-Cy5.5, PE-Cy7, Red laser fluorochromes are APC & APC-Cy7, Violet laser fluorochromes, BV421, BV511, BV570, BV605, BV650 & BV780. 22 bit and 7 log decade fluorescence allows PMT voltages to be pre-set making multi-colour analysis easier. There is an auto-sampler that allows automated analysis of 5 ml FACS tubes, 24, 48 and 96 well TC plates with washing and mixing between wells to reduce carry over. The syringe driven sampling system allows the automated calculation of absolute counts.
Cytek Aurora 5 laser, UV, Violet, Blue, YG and Red lasers 40+ fluorochrome flow cytometric spectral analyser with 96 well plate reader

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5042924/

Publications

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition). Klionsky DJ, Warnes G, et al Autophagy, 2020
G Warnes. Flow cytometric detection of zombie Jurkat cells, 2020. (manuscript in preparation).
HW King, N Orban, JC Riches, AJ Clear, G Warnes, SA Teichmann, L James. Antibody repertoire and gene expression dynamics of diverse human B cell states during affinity maturation. https://www.biorxiv.org/doi.org/10.1101/2020.04.28.054775, 2020.s, 1, 134-135, 56-66, 2018.
S Hannan, A Affandi, M Minere, C Jones, P Goh, B Popp, G Warnes, R Trollmann, D Nizetic, T Smart. Differential co-assembly of α1-GABAARs associated with epileptic encephalopathy, J Neuroscience, DOI: 10.1523/JNEUROSCI.2748-19.2020, 2020.
G Warnes. Flow cytometric detection of hyper‑polarized mitochondria
in regulated and accidental cell death processes, Apoptosis, Aug;25(7-8):548-557. doi: 10.1007/s10495-020-01613-5, 2020.
G Warnes. Flow cytometric detection of hyper‑polarized mitochondria in regulated and accidental cell death processes, Apoptosis, doi.org/10.1007/s10495-020-01613-5, 202
G Warnes. Flow Cytometry:- A new vista in cell death methodologies, Journal of Immunology Research and Therapy, 5(1): 212-214, (ISSN: 2472-727X), 2020.
G Warnes. Flow cytometric characterisation of accidental cell death highlights connections to regulated cell death, Journal of Cellular Signaling, 1(1):1-3, 2020.
A Vossenkamper, G Warnes.Thapsigargin induced apoptosis occurs with a delayed DNA damage response, Br J Cancer Res 2(4):314-319, 2019.
J Gibbons, K Marno, R Pike, WY Lee, C Jones, B Ogunkolade, C Pardieu, A Bryan, R Fu, G Warnes, P Rowley, R Sloan, Á McKnight. HIV-1 accessory protein Vpr interacts with REAF/RPRD2 to mitigate its antiviral activity, Journal of Virology, Nov 27. pii: JVI.01591-19. doi: 10.1128/JVI.01591-19. [Epub ahead of print], 2019.
Li X, Ahmad US, Huang Y, Uttagomol J, Rehman A, Zhou K, Warnes G, McArthur S, Parkinson EK, Wan H. Desmoglein-3 acts as a pro-survival protein by suppressing reactive oxygen species and doming whilst augmenting the tight junctions in MDCK cells, Mechanisms of Ageing and Development, Oct 30:111174. doi: 10.1016/ j.mad. 2019. 111174, 2019.
H Wan, A Rehman, Y Cai, C Hünefeld, H Jedličková, Y Huang, MT Teh, U Ahmad, J Uttagomol, A Kang, G Warnes, C Harwood, D Bergamaschi, KE Parkinson, M Röcken. The desmosomal cadherin desmoglein-3 acts as an anti-stress protein via suppression p53 in keratinocytes, J Cell Death & Disease, 5(10)10/750, 2019.
A Cossarizza, G Warnes et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies, Eur J Immunol. 2019 Oct;49(10):1457-1973. doi: 10.1002/eji.201970107, 2019.
A Vossenkamper, G Warnes. Flow cytometry reveals the nature of oncotic cells, Int. J. Mol. Sci. 20(18), 4379; 2019.
G Warnes. Determining the fate of cells with flow cytometry, Blog by Global-Engage.com, Flow Cytometry Congress, London, 2019.
G Warnes. Phenotypic flow cytometric analysis of regulated cell death - 124 sub-populations identified explain why flow cytometry is better here than Western Blot. ACEA Biosciences Webinar, 2019.
D Bergamaschi, A Vossenkaemper, WYJ Lee, P Wang, E Bochukova, G Warnes. Simultaneous polychromatic flow cytometric detection of multiple forms of regulated cell death. Apoptosis, 24(5) 453-464, 2019.
A Popat, AA Patel, G Warnes. A flow cytometric study of ER Stress and autophagy, Cytometry A, 95(6), 672-682, DOI: 10.1002/cyto.a.23665, 2019.
A Vossenkamper, G Warnes. A flow cytometric immunophenotyping approach to the detection of regulated cell death processes. Journal of Immunological Sciences, 2 (5) 2018.
HL Lee, R Pike, MHA Chong, A Vossenkamper, G Warnes. Simultaneous flow cytometric immunophenotyping of necroptosis, apoptosis and RIP1 dependent apoptosis Methods, 1, 134-135, 56-66, 2018.
S Sisay, L Lopez-Lozano, M Mickunas, A Quiroga-Fernandez, J Palace, G Warnes, R Alvarez-Lafuente, P Dua, U Meir. Untreated relapsing remitting multiple sclerosis patients show antibody production against Latent Epstein Barr Virus (EBV) antigens mainly in the periphery and innate immune IL-8 responses preferentially in the CNS. Journal of Neuroimmunology, 306, 40-45, 2017.
E Pantazi, E Gemenetzidis, MT Teh, SV Reddy, G Warnes, C Evagora, G Trigiante, MP Philpot. GLI2 is a novel regulator of β-catenin and is associated with loss of E-cadherin, cell invasiveness and long-term epidermal regeneration. Journal of Investigative Dermatology, Mar 11. pii: S0022-202X(17)31218-6. doi: 10.1016/j.jid.2016.11.046. [Epub ahead of print] 2017.
AA Patel, G Warnes. Cell cycle analyshttps://www.scientificarchives.com/journal/journal-of-cellular-signalingis of ER stress and autophagy, Autophagy, Ed N Gorbunov, INTECH pub, ISBN 978-953-51-4815-9, 2016.
ESICM LIVES 2016: part one : Milan, Italy. 1-5 October 2016. DJ Klionsky, G Warnes et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Intensive Care Med Exp. Sep;4(Suppl 1):27. doi: 10.1186/s40635-016-0098-x. 2016.
G Warnes. The multiplexing of assays for the measurement of early stages of apoptosis by polychromatic flow cytometry. Flow Cytometry- Recent Prospectives, Ed I Schmid, INTECH pub, ISBN 978-953-51-4346-8, 2016.
G Warnes.Flow cytometric measurement of autophagy: Get yourself to eat your way out of cancer and neurodegenerative diseases. Science Atlas Feb, 2016.
DJ Klionsky, G Warnes et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd Edition). Autophagy Jan 2 12(1), 1-222, 2016.
HDT Torrance, K Brohi, G Warnes, HC Owens, CJ Hinds, DJ Pennington, MJ O'Dwyer. T helper cell polarization following severe polytrauma. Intensive Care Medicine Experimental, 3 (Suppl 1) A848 DOI.10.1186/2197-425X-51-A848,2015.
G Warnes. Flow cytometric assays for the study of autophagy. Methods, doi:10.1016/j.ymeth.2015.03.027, 2015.
A Chikh, P Sanza, C Raimondi, O Akinduro, G Warnes, G Chiorino, C Byrne, CA Harwood, and D Bergamaschi. IASPP is a novel autophagy inhibitor in keratinocytes. Journal of Cell Science Apr28, 2014.
G Warnes. Measurement of autophagy by flow cytometry, Studies of Cell Function. Curr Protoc Cytom. Apr 1;68:9.45.1-9.45.10, 2014.
P Dua, G Warnes, A Chruch, D Martino, D Baker, G Giovannoni, UC Meier. Functional effects of anti-neuronal antibodies from patients with Tourette's syndrome and disorders associated with streptococcal infection. Neurology Psychiatry and Brain Research; 20(1):17–18 2014.
TS MacFie, R Poulsom, A Parker, G Warnes, T Boitsova, A Nijhuis, N Suraweera, APoehlmann, J Szary, R Feakins, R Jeffery, RW Harper, AM Jubb, JO Lindsay, A Silver. DUOX2 and DUOXA2 form the predominant enzyme system capable of producing the reactive oxygen species H2O2 in active ulcerative colitis and are modulated by 5-aminosalicylic acid. Inflammatory Bowel Disease, Jan 2014.
S Chikte, N Panchal, G Warnes. Use of LysoTracker dyes: A flow cytometric study of autophagy. Cytometry A Feb, 85(2), 169-178, 2014.
E Pantazi, E Gemenetzidis, G Trigiante, G Warnes, L Shan, X Mao, M Ikram, MT Teh, YJ Lu, M Philpott.GLI2 induces genomic instability in human keratinocytes by inhibiting apoptosis. Cell Death Dis. Jan 30.5:e1028 doi 10.1038/cddis 2013.535 2014.
E Karanasios, A Englezou, S Cohen, G Warnes. Analysis of Autophagy Regulation: Discussion of recent research and new technologies. Meeting report of the flow cytometry Euroscicon meeting in UK 3rd October 2013. Kindle Edition, Amazon, Honnao Publishing, Jan 2014.
Helgason V. (Meeting Chaired by G Warnes). Autophagy regulation: recent reasearch and new technologies. The Biomedical Scientist 57(12):722-724, 2013.
Luo J, Lee S, Wu D, Yeh J, Ellamushi H, Wheeler AP, Warnes G, Zhang Y, Bo X. P2X7 purinoceptors contribute to the death of Schwann cells transplanted into the spinal cord. Cell Death Dis. Oct 3;4:e829. doi: 10.1038/cddis.2013.343, Oct 2013.
A Want, S Cohen, G Warnes. Cell suicide is not painless for the flow cytometrist: New Assays to measure cell death. Meeting report of the flow cytometry Euroscicon meeting in UK 7th November 2012. Kindle Edition, Amazon, Honnao Publishing, June 2013.
N Panchal, S Chikte, BR Wilbourn, UC Meier, G Warnes. Flow cytometric measurement of cell organelle autophagy. Autophagy - A Double-Edged Sword - Cell Survival or Death? Ed, Y. Bailly. InTech Open Access Publisher ISBN 978-953-51-1062-0, April 2013.
RN Matin, A Chikte, SLP Chong, D Mesher, M Graf, P Sanza, V Senatore, M Scatolini, F Moretti, IM Leigh, CM Proby, A Costanzo, G Chiorino, R Cerio, CA Harwood, D Bergamaschi. p63 is an alternative p53 repressor in melanoma that confers chemoresistance and a poor prognosis. J Exp Med Feb, 210 (3), 581-603, 2013.
C Raimondi, A Chikh, AP Wheeler, T Maffucci, M Falasca. A novel regulatory mechanism links PLCg1 to PDK1. J Cell Sci Epub May18 2012.
KD Henseleit, AP Wheeler, G Warnes, KM Braun. Complementary Whole mount Visualization and Flow Cytometry Strategies. Progenitor Cells : Methods in Molecular Biology Vol 916 (3) p243-261. Eds KA Mace, KM Braun. ISBN 978-1-61779-979-2, 2012.
A Rosello, G Warnes, UC Meier. Cell death pathways and autophagy in the central nervous system and its involvement in neurodegeneration, immunity and CNS infection: to die or not to die - that is the question. Clinical & Experimental Immunology Apr, 162(1), 52-7, 2012.
DD Ateh, VH Leinster, SR Lambert, A Shah, A Khan, H Walklin, J Johnstone, N Ibrahim, M Kadam, Z Malik, M Girones, G Veldhuis, G Warnes, S Marino, I McNeish, JE Martins. The intracellular uptake of CD95 modified paclitaxel-loaded poly(lactic-co-glycolic acid) microparticles Biomaterials 32 (33), 8538-47, 2011.
SN Jayasinghe, G Warnes, CJ Scotton. Bio-electrosprayed living microenvironments implanted into mouse models. Macromolecular Bioscience 11(10), 1364-69, 2011.
G Warnes, S Martins. Real-time flow cytometry for the kinetic analysis of oncosis. Cytometry 79A, 181-191, 2011.
A. Tarnok. Review of Real-time flow cytometry for the kinetic analysis of oncosis. Cytometry 79A, 177-178, 2011.
R.Challis, G Warnes. Flow cytometry techniques and instrumentation: keeping up with the changes. The Biomedical Scientist 55 (2) : 94-96, 2011.
A Vossenkamper, O Marches, PD Fairclough, G Warnes, AJ Stagg, JO Lindsay, PC Evans,LA Luong, NM Croft, S Naik, G Frankel, TT McDonald. Inhibition of NF-kB signaling in human dendritic cells the enterpathogenic Escherichia coli effector protein NIeE . J Immunol 185: 4118-4127, 2010.
MG Valorani, A Germani, WR Otto, L Harper, A Biddle, CP Khoo, WR Lin, MI Hawa, P Tropel, MP Patrizi, P Pozzilli, MR Alison. Hypoxia increases Sca-1/CD44 co-expression in murine mesenchymal stem cells and enhances their adipogenic differentiation potential. Cell Tissue Res, DOI 10.1007/s00441-010-0982-8 2010.
G Warnes. Flow Cytometric Applications of FRET - Fluorescence Resonance Energy Transfer. The Biomedical Scientist, 53(2) p119-121,Feb 2009.
CP Khoo, MG Valorani, M Brittan, MR Alison, G Warnes, U Johansson, M Hawa, P Pozzilli. Characterization of endothelial progenitor cells in the NOD mouse as a source for cell therapies. Diabetes Metab Res Rev 25(1): 89-93, 2009.
SM Janes, TA Ofstad, DH Campbell, A Eddaoudi, G Warnes, D Davies, F Watt. PI3-kinase dependent activation of apoptotic machinery occurs on commitment of epidermal keratinocytes to terminal differentiation. Cell Res 19(3): 328-339, 2009.
JP Biggerstaff, B Weidow, J Dexheimer, G Warnes, J Vidosh, S Patel, M Newman, P Patel. Soluble fibrin inhibits lymphocyte adherence and cytotoxicity against tumor cells:Implications for cancer metastasis and immunotherapy. Clin Appl Thromb Hemost 14(2): 193-202, 2008.
P Takousis, P Johonnett, J Williamson, P Sasieni, G Warnes, T Forshew, V Azuara, A Fisher, PJ Wu, PJ Jones, R Vatcheva, S Beck, D Sheer. Replication timing profile reflects the distinct functional and genomic features of the MHC Class II Region. Cell Cycle Oct 6:19, 2393-2398, 2007.
S Leverrier, D Bergamaschi, L Ghali, A Ola, G Warnes, B Akgul, K Blight, R Garcia-Escudero, A Penna, A Eddaoudi, A Storey. Role of HPV E6 proteins in preventing UVB-induced release of pro-apoptotic factors from the mitochondria. Apoptosis 12 (3), 549-60, 2007.
G Warnes, MG Macey. Calcium: Cytoplasmic, mitochondria, endo-plasmic reticulum and flux measurements, Flow Cytometry: Principles & Applications. Ed MG Macey, Springer, Humana Press ISBN 978-1-58829-691-7, 2007.
V Azuara, P Perry, S Sauer, M Spivakov, HF Jorgensen, RM John, M Gouti, M Casanova, G Warnes, M Merkenschlager, AG Fisher. Chromatin signatures of pluripotent cell lines. Nature Cell Biol. 8, 532-538, 2006.
P Perry, S Saur, N Billion, WD Richardson, M Spivakov, G Warnes, FJ Livesey, M Merkenschlager, AG Fisher, V Auara. A dynamic switch in the replication timing of key regulator genes in embryonic stem cells upon neural induction. Cell Cycle 3, (12), 1645-50, 2004.
MR Ehrenstein, JG Evans, A Singh, S Moore, G Warnes, DA Isenberg, C Mauri. Compromised function of regulatory T cells in Rheumatoid Arthritis and reversal by anti-TNF therapy. J Exp Med. 200, 277-285, 2004.
P Sabbattini, M Lundgren, A Georgiou, C Chow, G Warnes, N Dillion. Binding of Ikaros to the lambda5 promoter silences transcription through a mechanism that does not require heterochromatin formation. EMBO J. 20(11), 2812-22, 2001.
PH Dixon, et al, G Warnes, C Williamson. Heterozygous MDR3 missense mutation associated with intrahepatic cholestasis of pregnancy: evidence for a defect in protein trafficking. Hum Mol Genet. 9(8), 1209-17, 2000.
G Warnes, JP Biggerstaff, JL Francis. Monocyte B7 and Sialyl Lewis X modulates the efficacy of IL-10 down-regulation of LPS-induced monocyte tissue factor in whole blood. British Journal of Haematology 102,588-596, 1998.
VA Gant, G Warnes, I Phillips, GF Savidge. The application of flow cytometry to the study of bacterial responses to antibiotics. J Med Microbiol 39,1475-154, 1993.

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