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Flow Cytometry


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Overview

The Flow Cytometry Facility at Barts Cancer Institute provides a state of the art facility to support our research, 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 labeled probes we can perform:

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

→ Find out about the 'Google Earth of Cancer' CRUK Grand Challenge Project


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Using the Facility

Internal Users can find joining instructions, pricing and protocols on the Intranet.

Cell sorting and CyTOF2 must be arranged directly with the flow cytometry core staff using flowcytometry@qmul.ac.uk

External Users are welcome, particularly for ImageStream and CyTOF2.  Please contact the Facility Manager
Becki Pike (r.pike@qmul.ac.uk) Tel: 020 7882 2119

 

Flow Cytometry Analysers

Analyser Fortessa

The following Analysers are installed in the BCI Flow Cytometry Facility and are available for independent use:

BD LSRFortessa 1
Lasers: Blue, Red, Violet, Yellow-Green
(16 fluorescent parameters)
Build a Panel with Fluorofinder here


BD LSRFortessa 2
Lasers: Blue, Red, Violet, Yellow-Green
(16 fluorescent parameters)
Build a Panel with Fluorofinder here


BD LSRFortessa 3
Lasers: Blue, Red, Violet
(14 fluorescent parameters)
Build a Panel with Fluorofinder here


Analyser CaliburBD FACSCalibur
Lasers: Blue, Red
(4 fluorescent parameters)
Build a Panel with Fluorofinder here


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 by flow facility staff. Please contact us directly at flowcytometry@qmul.ac.uk.


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BD FACSAria II

Lasers: Blue, Red, Violet, Yellow-Green
(14 fluorescent parameters)

Build a Panel with Fluorofinder here


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BD FACSAria Fusion

Lasers: Blue, Red, Violet 
(14 fluorescent parameters)

Build a Panel with Fluorofinder here

 

ImageStream X MkII

ImageStream

As well as generating conventional flow cytometry data, the Imagestream provides brightfield and fluorescence microscopy 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 here

 

CyTOF2

CyTOFimg2The 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 considerably 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 flowcytometry@qmul.ac.uk

External users are welcome - please contact the Facility Manager for information (contact details above).

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.


Analysis

Facility Acknowledgement

Please ensure that you acknowledge the BCI 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 BCI 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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Contact us

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For more information about the service please contact

Dr Becki Pike - Flow Cytometry Service Manager

Coming Soon – Helios & Hyperion Tissue Imager

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We are excited to announce that the 3rd generation CyTOF instrument – Helios – and the officially unreleased Hyperion Tissue Imager will be available soon.

Hyperion is an incredibly powerful imaging tool allowing labelling of tissue sections with up to 37 different antibodies.
More information

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