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 Medical and Dental Sciences (MDS), 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 Specialist
Dr Guillaume Desanti
Email: g.desanti@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
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
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.
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 request 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 Fluidigm 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
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. We will also require a valid grant code for recharge. Please provide it in the following structure:
- Organisation Unit Example: C082 (This will refer to your Institute)
- Finance Project Code Example 19779 (This should be a 5-7 digits number)
Account requests and flow cytometry training requests may be submitted simultaneously. New GCP level users are requested to submit a new project form through Stratocore before requesting flow cytometry training.
Analysers
BD LSRFORTESSA X-20
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.
| Fortessa 1, 2 & 3 |
|---|
|
Laser
|
Filter
|
Suggested Dye(s)
|
|
405 (Violet)
|
450/50
|
Pacific Blue,
BV450, CFP, Dapi
|
|
|
525/50
|
Pacific Orange,
BV510, AmCyan
|
|
|
610/20
|
BV605
|
|
|
670/30
|
BV650
|
|
|
710/50
|
BV711
|
|
|
780/60
|
BV786
|
|
488 (Blue)
|
530/30
|
FITC, GFP
|
|
|
670/30
|
PerCP, PerCPCy5.5
|
|
561 (Yellow)
|
586/15
|
PE, DsRed
|
|
|
610/20
|
PE-TxRed, mCherry, RFP, PI
|
|
|
670/30
|
PE-Cy5
|
|
|
710/50
|
PE-Cy5.5
|
|
|
795/71
|
PE-Cy7
|
|
642 (Red)
|
671/30
|
APC, AF647
|
|
|
722/44
|
AF700, Dylight680
|
|
|
795/70
|
APC-Cy7
|
| Fortessa 4 |
|---|
|
Laser
|
Filter
|
Suggested Dye(s)
|
|
355 (UV)
|
740/35
|
BUV737
|
|
|
379/28
|
BUV395
|
|
405 (Violet)
|
450/50
|
Pacific Blue, BV450, BV421, CFP, Dapi
|
|
|
525/50
|
Pacific Orange, BV510, AmCyan
|
|
|
610/20
|
BV605
|
|
|
670/30
|
BV650
|
|
|
710/50
|
BV711
|
|
|
780/60
|
BV786
|
|
488 (Blue)
|
530/30
|
FITC, GFP
|
|
|
670/30
|
PerCP, PerCPCy5.5
|
|
561 (Yellow)
|
586/15
|
PE, DsRed
|
|
|
610/20
|
PE-TxRed, mCherry, RFP, PI
|
|
|
670/30
|
PE-Cy5
|
|
|
710/50
|
PE-Cy5.5
|
|
|
795/71
|
PE-Cy7
|
|
642 (Red)
|
671/30
|
APC, AF647
|
|
|
722/44
|
AF700, Dylight680
|
|
|
795/70
|
APC-Cy7
|
Cytoflex
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 acquisition software is CytExpert.
| Cytoflex |
|---|
|
Laser
|
Filter
|
Suggested Dye(s)
|
|
405 (Violet)
|
450/50
|
Pacific Blue, BV450, eFluor450, BV421
|
|
|
525/40
|
Pacific Orange, BV510, AmCyan
|
|
|
610/20
|
BV605, Qdot605, Violet610
|
|
|
660/10
|
BV650, Qdot695, Violet660
|
|
610 (Yellow)
|
610/20
|
ECD, mCherry
|
|
|
585/42
|
PE, DsRed
|
|
|
390/50
|
PECy5.5, PI
|
|
|
780/60
|
PeCy7
|
|
638 (Red)
|
660/20
|
APC, AF647, eFlour660
|
|
|
712/25
|
AF700, Alexa Fluor 700, Dylight680
|
|
|
780/60
|
APC eFluor780, APC-Cy7, APC-A750
|
|
488 (Blue)
|
525/40
|
FITC, GFP, Alexa 488, CFSE
|
|
|
585/42
|
PercP
|
Attune NxT
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.
| Attune NxT |
|---|
|
Laser
|
Filter
|
Suggested Dye(s)
|
|
585 (Yellow)
|
YL1 585/16
|
PE, Alexa Fluor 568, PI, DsRed
|
|
|
YL2 620/15
|
PE-Texas Red, mCherry
|
|
|
YL3 695/40
|
PECy5.5, Alexa Fluor 700, PercP Cy5.5
|
|
|
YL4 780/60
|
PeCy7, Qdot800
|
|
488 (Blue)
|
BL1 530/30
|
FITC, GFP, Alexa 488, CFSE
|
|
|
BL2 590/40
|
PI, PE-Texas Red, PE-Alexa Fluor 610
|
|
|
BL3 695/40
|
PercP, PECy5.5, PercP Cy5.5
|
Cell Sorter
BD FACSAria™ Fusion
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.
| FACSAria |
|---|
|
Laser
|
Filter
|
Suggested Dye(s)
|
|
355 (UV)
|
450/50
|
DAPI, Hoechst Blue
|
ARIA 1
|
|
|
670/25
|
BUV661
|
ARIA 1
|
|
|
379/28
|
BUV395
|
ARIA 2
|
|
|
740/35
|
BUV737
|
ARIA 2
|
|
405 (Violet)
|
450/50
|
Pacific Blue, BV450, BV421, CFP, Dapi
|
|
|
525/50
|
Pacific Orange, BV510, AmCyan
|
|
|
610/20
|
BV605
|
|
|
660/20
|
BV650
|
|
|
710/50
|
BV711
|
|
|
780/60
|
BV786
|
|
488 (Blue)
|
695/40
|
PerCPCy5.5
|
|
|
530/30
|
FITC
|
|
561 (Yellow/Greeen)
|
582/15
|
PE, DsRed
|
|
|
610/20
|
PE-TxRed, mCherry, RFP, PI
|
|
|
670/14
|
PE-Cy5
|
|
|
710/50
|
PE-Cy5.5
|
|
|
780/60
|
PE-Cy7
|
|
640 (Red)
|
780/60
|
APC-Cy7
|
|
|
733/45
|
AF700, Dylight680
|
|
|
670/30
|
APC
|
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.
The UoB has invested and supports state of the art flow cytometry, sorting and mass cytometry equipment. It is important for the facility to be properly recognised and its outputs measured. This is to ensure that this facility continues to be supported by the UoB to allow for the maintenance of existing equipment and to expand the facility with new hardware as appropriate. We therefore request that all users of the facility recognise its contribution to manuscripts. You can use the following statement for publications.
The authors would like to acknowledge the Flow Cytometry Platform at the University of Birmingham for support of flow/mass cytometry experiments. The authors would like to acknowledge (flow cytometry or mass cytometry specialist name) for advice with design acquisition of flow cytometry or sorting experiments.
- 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.