Blizard Institute - Barts and The London

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 four laser 14 fluorochrome analyser a BD LSRII, a three laser 8 fluorochrome BD FACSCanto II and a 3 laser 13 fluorochrome ACEA Biosciences Novocyte 3000.

SPADE software (fluidigm inc), a year subsrciption was purchased by Themes funding for Users to allow multi-dimensional analysis of FACS data by Spanning Tree Progression of Density Normalized Events or SPADE, see the link https://fluidigm.cytobank.org/cytobank/ please enquire for login details, see the link for the capabilities of the software, pdf. The software allows the automated identification of subpopulations of cell samples labelled with up to 40 fluorophores.

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 Dr Dan Pennington, Immunobiology Centre Lead.

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

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

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 - Four laser 14 colour flow cytometric analyser with Blue laser which detects FITC, PE, PE-CF595, PerCP or PE-Cy5, PerCP-Cy5.5 pr PE-Cy5.5 & PE-Cy7, Red laser detects APC, AF700 & APC-Cy7, Violet laser detects BV421, BV511, BV605, UV laser detects violet and green Indo-1, DAPI or 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.
  • SFIF funding will allow the purchase of a 5 laser, 40+ fluorochrome flow cytometric analyser - coming in Summer 2020 

 

 

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