Flow Cytometry

The Flow Cytometry Facility at the William Harvey Research Institute provides a state of the art facility to support the important research being carried out at the Institute, within Queen Mary University of London. As well as a number of flow cytometry analysers and cell sorters, the facility also houses an ImageStream X Mk II imaging cytometer and CyTOF2 mass cytometer.

Flow Cytometry is a versatile technique enabling high throughput analysis and cell separation. Using fluorescent proteins, dyes, and fluorescently labelled probes we can perform:

  • cellular phenotyping
  • functional assays
  • cell cycle analysis
  • intracellular protein studies e.g. cytokines and transcription factors
  • RNA quantification, and more.  

Training

All internal users require an iLab account with associated budget code. An account can be created at:
https://qml.corefacilities.org/account/signup/596

Once this is in place, please contact the facility staff for an induction and training. Unsupervised use will only be permitted after the user can demonstrate their ability to use the machine safely.

Booking

Analysers and Image Stream are bookable using the iLab system.

Cell sorting and CyTOF2 must be arranged directly with the Flow Cytometry core staff. Please contact This email address is being protected from spambots. You need JavaScript enabled to view it.

External users are welcome, particularly for ImageStream and CyTOF2. Please contact the Facility Manager Becki Pike (This email address is being protected from spambots. You need JavaScript enabled to view it.) Tel: 020 7882 2119.

Location: JVSC 2nd floor, adjacent to Clinical Pharmacology

Flow Cytometry Analysers

The following analysers are installed in the flow lab and are available for independent use:

BD LSRFortessa 1 - Lasers: Blue, Red, Violet, Yellow-Green (16 fluorescent parameters)
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BD LSRFortessa 2 - Lasers: Blue, Red, Violet, Yellow-Green (16 fluorescent parameters)
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BD LSRFortessa 3 - Lasers: Blue, Red, Violet (14 fluorescent parameters)
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BD FACSCalibur - Lasers: Blue, Red (4 fluorescent parameters)
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Cell Sorting

We have two BD Aria Cell Sorters with capacity for up to 4-way cell sorting, as well as single cell sorting into 96- and 384-well plates. Cell sorting is run as a service for researchers by flow facility staff. Please contact us directly at This email address is being protected from spambots. You need JavaScript enabled to view it.  

BD FACSAria II
Lasers: Blue, Red, Violet, Yellow-Green
(14 fluorescent parameters available) 

Build a Panel with Fluorofinder here

 BD FACSAria Fusion
Lasers: Blue, Red, Violet
(14 fluorescent parameters available) 

Build a Panel with Fluorofinder here 

Aria II Aria Fusion

IMAGESTREAM X MkII

As well as generating conventional flow cytometry data, the Imagestream provides brightfield and fluorescence images of each cell at 20x, 40x or 60x magnification. This is a 3 laser instrument (red, blue and violet) which can be used to look at up to 10 different fluorescent parameters - ideal for investigating co-localisation, internalisation, cell mitosis, immunological synapse and much more.

Build a Panel with Fluorofinder 

CyTOF2 Mass Cytometry


The CyTOF2 provides next generation single cell analysis, with the ability to investigate up to 50 parameters in one tube. Mass Cytometry labels cells with antibodies conjugated to stable heavy metal isotopes in the place of fluorochromes. These probes produce signal in only one mass channel, avoiding the problem of spectral overlap experienced in flow cytometry, enabling investigation of considerable more parameters per single sample. This makes it the perfect tool for gaining a large amount of information where sampling material is limited, and for carrying out complex phenotyping of rare cell types.

CyTOF2 is run as a service for researchers by facility staff. Please contact directly at This email address is being protected from spambots. You need JavaScript enabled to view it..

External users are welcomePlease contact the Facility Manager Becki Pike (This email address is being protected from spambots. You need JavaScript enabled to view it.) Tel: 020 7882 2119.

Analysis

FlowJo, BD FACSDiva, and CellQuest Pro dongles are available for offline data analysis. A number of computers are also available for offline analysis with BD FACSDiva, FlowJo and IDEAS (for ImageStream data).

Core staff can also provide training and assistance with the above software.

Facility Acknowledgement

Please ensure that you acknowledge the WHRI Flow Cytometry Facility in any publications or presentations that include work carried out in the facility. This is vital to ensure continued funding for the facility, as the cost recovery model in place only partly covers total running costs.

A brief sentence in the acknowledgement section of a paper is sufficient:

“This research was supported by the WHRI Flow Cytometry Facility.”

You may also wish to acknowledge an individual member of staff (delete as appropriate):

“In particular, we thank [name] for their advice and support in flow cytometry/cell sorting/CyTOF/ImageStream”

Thank you in advance for your support.

 

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