The Flow Cytometry platform at the University of Birmingham supports flow cytometry, cell sorting and mass cytometry across the University of Birmingham (UoB); the Technology Hub Flow Cytometry core facility is based in the Institute of Biomedical Research (IBR) in the College of Medicine and Health (CMH), the Flow Cytometry team also support instruments in the UoB Laboratories at the Queen Elizabeth Hospital, the Institute of Translational Medicine and the College of Life and Environmental Sciences (LES).
The Flow Cytometry facility has undergone recent expansion with a broad portfolio of state-of-the-art flow cytometers and the Helios (CyTOF) mass cytometer. It provides a range of services from user training on standard self-service instruments, through to a bespoke service of cell sorting and mass cytometry. The platform has recently expanded into the delivery of GCP level service, and into mass cytometry in our dedicated GCP level laboratories within the ITM. The Flow Cytometry Unit is accessible to all UoB current students and staff, all UK academics and commercial organisations.
Flow Cytometry Team
Flow Cytometry Specialist
Dr Guillaume Desanti
Guillaume joined the Flow Cytometry facility at the end of 2021. He has been awarded a PhD in Immunology (2007), a Master in Immunology (2003) and a Bachelor in Biochemistry by Paris7/Denis Diderot University (France). Over his nineteen years of doctoral and postdoctoral experience, Guillaume used flow cytometry in the research fields of host and pathogen interactions, autoimmune diseases, inflammation, lymphopoiesis and lymphoid tissue development. He has experience in a range of flow cytometry equipment including Beckman Cyan (analyser), Beckman MoFlo Astrios EQ (cell sorter), BD Fortessa X-20 (analyser), BD FACS Aria Fusion (cell sorter) and ThermoFisher Attune NxT (analyser).
Email: g.desanti@bham.ac.uk
Flow Cytometry Specialist
Ferdus Sheik
Ferdus joined the Flow Cytometry facility in 2022. He has been awarded an BSc in Applied Biomedical Science (2017) from Aston University and an MSc in Clinical Haematology and Transfusion Science (2021) from Manchester Metropolitan University. Originally from a clinical background, Ferdus has gained experience of flow cytometry through the completion of the Scientist Training Programme (STP) in which he established a ‘multi-parameter minimal residual disease panel to monitor chronic lymphocytic leukaemia in patients’ and aided in diagnostic immunophenotyping of haematological malignancies at Nottingham University Hospitals Trust. He has experience in a range of flow cytometry equipment including Beckman Coulter Navios and BD FACSLyric.
Email: f.sheik@bham.ac.uk
Mass Cytometry Specialist
Dr Shahram Golbabapour
Shahram joined the Flow Cytometry facility in 2019. He was awarded a PhD in Biotechnology and Biochemistry (2015) from the University of Malaya (Malaysia), a Masters in Biotechnology, and Doctor of Veterinary Medicine degree (DVM). Shahram has four years postdoctoral experience and advanced knowledge of a range of flow cytometry equipment (sorter and analysis) including Beckman MoFlo Astrios EQ (sorter), BD FACS Aria (sorter), BD Celesta (analysis), BD Fortessa (analysis) the Helios (CyTOF) Mass Cytometer.
Email: S.Golbabapour@bham.ac.uk
Research Facilities Manager
Dr Adriana Flores-Langarica
Email: A.FloresLangarica@bham.ac.uk
Financial Assistant
James Batten
Email: j.batten@bham.ac.uk
Flow Cytometry Academic Lead
Dr Kai-Michael Toellner
Email: K.M.TOELLNER@bham.ac.uk
Deputy Flow Cytometry Academic Lead
Dr William Jenkinson
Email: W.E.Jenkinson@bham.ac.uk
Training and Support
We provide training and support for users to independently use all our available analysers. Currently our training programmes are divided in two sections. Firstly, the CANVAS Module which can be completed at the user pace and once that is completed we deliver an in person training session at the equipment. If you require training just request it in Stratocore. We deliver sessions regularly.
Once trained, users will initially have ‘novice’ rights in which they can only book within peak hours. After 20 hours experience with the equipment, users will have ‘autonomous’ rights meaning they can book 24/7.
We also organize seminars relevant to our users. Those are advertised locally but you can also follow us on Facebook @ Flowcytometry Bham for further details and news about the Flow Cytometry Platform.
We deliver a full service for cell sorting. Additionally, to support users that their sorting needs are outside working hours we have a program of super-user training. All super-user requests should be submitted to flowcytometry@contacs.bham.ac.uk explaining your need for access after-hours access to the sorter and should include the relevant PI. All requests will be evaluated by our Operations Team.
For Mass Cytometry we deliver a full service where a specialist will start, QC and run your samples, so no training is required. However, we work closely with Standard BioTools to provide support to users from panel design, sample processing and analysis.
Finally, as part of the Midlands Innovation Flow Cytometry Group we deliver an Introduction to Flow Cytometry course aimed to people approaching the technology. The sessions are delivered online and include a theory and a live demonstration in a flow cytometer. These sessions take place once every quarter and are advertised widely.
Online support
Online Tools
Protocols
Access and Booking
External users are encouraged to contact us via email in the first instance, however all internal prospective users will need to request an account on the core facilities booking system Stratocore (unless you already have one). Please ensure you use your UoB email address to register your Stratocore account.
As part of your account, you will need a valid finance/grant code. We need this to be able to charge you for your use of the equipment.
You code will either be a project or general ledger. Guidance on finance/grant codes is here:
Project Ledger. (Project Finance Number begins with a ‘1’ or ‘2’). Project Ledger codes require 3 components in the format:
Cost Centre Code - Task Number - Project Finance Number
C082 - 1.1 - 21991
General Ledger. (Project Finance Number begins with a ‘6’). General Ledgers require 2 components in the format:
Cost Centre Code - Project Finance Number
C283 - 65371
If you are unsure about any of your Finance code, contact techubfinance@contacts.bham.ac.uk who will be able to assist you.
Account requests and flow cytometry training requests may be submitted simultaneously. GCP level users are required to submit a new project form through Stratocore before requesting flow cytometry training.
For any Stratocore or finance relation queries please contact techubfinance@contacts.bham.ac.uk
Equipment
Analysers
The BD LSRFortessa™ X-20 cell analyser delivers high-performance multicolour analysis. The analyser has 5 lasers to detect up to 16 parameters simultaneously to support ever increasing demands in multicolour flow cytometry. The acquisition software is DIVA.
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Fortessa 2 & 3
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Laser
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Filter
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Suggested Dye(s)
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405 (Violet)
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450/50
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Pacific Blue,
BV450, CFP, Dapi
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525/50
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Pacific Orange,
BV510, AmCyan
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610/20
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BV605
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670/30
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BV650
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710/50
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BV711
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780/60
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BV786
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488 (Blue)
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530/30
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FITC, GFP
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670/30
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PerCP, PerCPCy5.5
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561 (Yellow)
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586/15
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PE, DsRed
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610/20
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PE-TxRed, mCherry, RFP, PI
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670/30
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PE-Cy5
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710/50
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PE-Cy5.5
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795/71
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PE-Cy7
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642 (Red)
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671/30
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APC, AF647
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722/44
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AF700, Dylight680
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795/70
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APC-Cy7
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Fortessa 1 & 4
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Laser
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Filter
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Suggested Dye(s)
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355 (UV)
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740/35
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BUV737
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379/28
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BUV395
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405 (Violet)
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450/50
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Pacific Blue, BV450, BV421, CFP, Dapi
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525/50
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Pacific Orange, BV510, AmCyan
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610/20
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BV605
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670/30
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BV650
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710/50
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BV711
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780/60
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BV786
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488 (Blue)
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530/30
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FITC, GFP
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670/30
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PerCP, PerCPCy5.5
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561 (Yellow)
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586/15
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PE, DsRed
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610/20
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PE-TxRed, mCherry, RFP, PI
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670/30
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PE-Cy5
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710/50
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PE-Cy5.5
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795/71
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PE-Cy7
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642 (Red)
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671/30
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APC, AF647
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722/44
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AF700, Dylight680
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795/70
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APC-Cy7
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Beckman Coulter CytoFLEX S
The high-performance CytoFLEX system utilizes three excitation lines with independent, alignment-free focusing optics, simultaneous 13-parameters and two scatter parameters. It has superior sensitivity and resolution for excellent fluorescence and nanoparticle detection. The instrument also possesses a plate loader module, enabling the acquisition of samples within standard flat-bottom, U-bottom and V-bottom 96-well plates. The acquisition software is CytExpert.
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Beckman Coulter CytoFLEX S
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Laser
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Filter
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Suggested Dye(s)
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405 (Violet)
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450/50
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Pacific Blue, BV450, eFluor450, BV421
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525/40
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Pacific Orange, BV510, AmCyan
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610/20
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BV605, Qdot605, Violet610
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660/10
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BV650, Qdot695, Violet660
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610 (Yellow)
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610/20
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ECD, mCherry
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585/42
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PE, DsRed
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390/50
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PECy5.5, PI
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780/60
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PeCy7
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638 (Red)
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660/20
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APC, AF647, eFlour660
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712/25
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AF700, Alexa Fluor 700, Dylight680
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780/60
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APC eFluor780, APC-Cy7, APC-A750
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488 (Blue)
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525/40
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FITC, GFP, Alexa 488, CFSE
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585/42
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PercP
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The Attune NxT Flow Cytometer system makes flow cytometry available to both new and experienced researchers. It has 2 lasers and can detect 7 parameters and two scatter parameters. I also has available an auto sampler to acquire from 96 well plates. The acquisition software is Invitrogen Attune NxT.
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Attune NxT
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Laser
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Filter
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Suggested Dye(s)
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585 (Yellow)
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YL1 585/16
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PE, Alexa Fluor 568, PI, DsRed
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YL2 620/15
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PE-Texas Red, mCherry
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YL3 695/40
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PECy5.5, Alexa Fluor 700, PercP Cy5.5
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YL4 780/60
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PeCy7, Qdot800
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488 (Blue)
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BL1 530/30
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FITC, GFP, Alexa 488, CFSE
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BL2 590/40
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PI, PE-Texas Red, PE-Alexa Fluor 610
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BL3 695/40
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PercP, PECy5.5, PercP Cy5.5
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Imaging Flow Cytometer
The Amnis ImageStream Mk II is a 6-channel imaging flow cytometer which combines the speed and sensitivity of flow cytometry with the detailed imagery and functional insights of microscopy. By collecting large numbers of digital images per sample and providing a numerical representation of image-based features, the instrument combines the per cell information content provided by standard microscopy with the statistical significance afforded by large sample sizes common to standard flow cytometry. The acquisition software is INSPIRE whereas the analysis software is IDEAS.
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Amnis ImageStream Mk II Standard Detector Configuration
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Channel
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Filter (nm)
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Excitation Laser (nm)
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405
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488
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642
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785
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1
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435 – 505
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*DAPI, BV421, AF405,
Hoechst, PacBlue, CascadeBlue, eFluor450, DyLight405, CFP, Live/Dead Violet
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|
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2
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505 – 560
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*BV510, PacOrange, CascadeYellow, AF430, eFluor525, QD525
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FITC, AF488, GFP, YFP, DyLight488, PKH67, Syto13, SpectrumGreen, LysoTrackerGreen, MitoTrackerGreen
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3
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560 -595
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*QD565, QD585, eFluor565
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PE, PKH26, DSRed, mOrange, CellMask/CellTracker/SYTOX Orange, Cy3
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|
|
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4
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595 – 642
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*QD625, eFluor625, BV605
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PE-TexRed, ECD, PE-AF610, RFP, QD625, eFluor625
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|
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5
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642 – 745
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*QD705, eFluor700, BV711
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PE-Cy5, PE-AF647, 7AAD, PI, PerCP, PerCP-Cy5.5, DRAQ5, QD705, eFLuor650, FuraRedlo, DRAQ5, LDS 751
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APC, AF647, AF660, AF680, DRAQ5, Cy5, DyLight649, DyLight680, PE-AF647, PE-Cy5, PerCP, PerCP-Cy5.5
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6
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745-785
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*QD800, BV786
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PE-Cy7, PE-AF750, QD800
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APC-Cy7, APC-AF750, APC-H7, APC-eFluor750, Cy7, AF750, DyLight750, PE-Cy7, PE-AF750
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SSC
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Recommended dyes (based on optimal excitation and detection channels) are in boldface.
*Many dyes will excite by more than one laser, and this can increase cross camera compensation.
Ideal dyes are DAPI (or BV421), AF488, PE, PE-TxRed, AF647, SSC/BF
Cell Sorter
The BD FACSAria™ Fusion flow cytometer delivers exceptional multicolour performance. The fully integrated biosafety cabinet meets emerging operator and sample protection requirements as well as global standards for bio-protection. It has 5 lasers to detect up to 18 parameters simultaneously. It is capable of sorting samples in tubes or plates. With the capacity of 4 way sorting. The acquisition software is DIVA.
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FACSAria
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Laser
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Filter
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Suggested Dye(s)
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355 (UV)
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450/50
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DAPI, Hoechst Blue
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ARIA 1
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670/25
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BUV661
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ARIA 1
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379/28
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BUV395
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ARIA 2
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740/35
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BUV737
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ARIA 2
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405 (Violet)
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450/50
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Pacific Blue, BV450, BV421, CFP, Dapi
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|
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525/50
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Pacific Orange, BV510, AmCyan
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|
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610/20
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BV605
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|
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660/20
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BV650
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|
|
710/50
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BV711
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|
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780/60
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BV786
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|
488 (Blue)
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695/40
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PerCPCy5.5
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|
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530/30
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FITC
|
|
561 (Yellow/Greeen)
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582/15
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PE, DsRed
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|
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610/20
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PE-TxRed, mCherry, RFP, PI
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|
|
670/14
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PE-Cy5
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|
|
710/50
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PE-Cy5.5
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|
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780/60
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PE-Cy7
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|
640 (Red)
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780/60
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APC-Cy7
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|
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733/45
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AF700, Dylight680
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|
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670/30
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APC
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Mass Cytometer
Helios™ Mass Cytometer - CyTOF®
Helios™, a CyTOF® system, comprises the most recent advances in mass cytometry and is designed to provide a new and improved tool for bioanalytical single-cell detection and analysis. This high-performance mass cytometer, from the CyTOF family of instruments, enables the analysis of more than 40 markers and uniquely allows their quantitative determination with negligible spectral overlap, a result of exquisite resolution between mass detection channels.
Acknowledgement Statement
The University of Birmingham (UoB) is a founding signatory of ‘The Technician Commitment’. A sector wide, collaborative initiative that aims to ensure visibility, recognition, career development and sustainability for technical staff working in higher education and research, across all disciplines.
The UoB has invested and supports the Technology Hub facilities not only with equipment but also specialist staff. The staff are a key element of our offering since they not only ensure that all equipment is looked after and performing to specification, but they also have a significant experience, and the input and advice is very valuable to our users.
In line with this, the Technology Hub Facilities request that all users of the facilities to recognize its contribution to manuscripts in one of the following ways:
The authors would like to acknowledge the (Facility) at the UoB for support of (technology used) experiments. The authors would like to acknowledge (specialist name).
In some instances, the specialist may make substantial contributions to the conception / design of the work or the acquisition, analysis, or interpretation of data for a manuscript. In this case, it is expected that this is appropriately recognized through co-authorship on any publications resulting from the work. Criteria for inclusion of a specialist as an author on a publication. Where the specialist has either.
Performed experiments or analysis, the data from which is included in the manuscript.
Produced customized analysis workflows or scripts for the work.
Significantly contributed to the design, conception, or interpretation of the experiments.
Publications
- CD103+CD11b+ mucosal classical dendritic cells initiate long-term switched antibody responses to flagellin. Flores-Langarica A, Müller Luda K, Persson EK, et al. (2017). Mucosal Immunol. May;11(3):681-692. doi: 10.1038/mi.2017.105. Epub 2017 Dec 20.
- Intestinal CD103+CD11b+ cDC2 Conventional Dendritic Cells Are Required for Primary CD4+ T and B Cell Responses to Soluble Flagellin. Flores-Langarica Adriana, Cook Charlotte, Müller Luda Katarzyna, et al. (2019). Frontiers in Immunology. 9: 2409. doi: 10.3389/fimmu.2018.02409.
- Antigen and checkpoint receptor engagement recalibrates T cell receptor signal strength. Thomas, AE., Jennings, E., Lecky, DA., Thawait, N., Flores-Langarica, A., Copland, A., Maslowski, K., Wraith, DC & Bending D. (2021) Immunity. 13;S1074-7613(21)00352-6. doi: 10.1016/j.immuni.2021.08.020.
Contact us
flowcytometry@contacts.bham.ac.uk
cytof@contacts.bham.ac.uk
techubfinance@contacts.bham.ac.uk
Research Facilities Manager
Dr Adriana Flores-Langarica
Email: A.FloresLangarica@bham.ac.uk
Flow Cytometry Academic Lead
Dr Kai-Michael Toellner
Email: K.M.TOELLNER@bham.ac.uk