Flow Cytometry Facility

The Sir William Dunn School of Pathology Flow Cytometry Facility is open to scientists from the University of Oxford as well as external users. We pride ourselves on offering the best service we can from the start to finish of any experiment involving flow cytometry. This includes advice on cell nomenclature, sample preparation, panel design, compensation of fluorescent spill over, flow data analysis, and cell sorting.

Flow Cytometry is a powerful technique that permits the analysis of millions of cells in a matter of minutes at a single cell resolution. The main advantages of this technique are accuracy and speed. Typical applications include: immunophenotyping, cell-cycle analysis, proliferation analysis, and analysis of transfection efficiency. This is achieved through analysis of fluorescent tags conjugated to surface or intracellular domain of cells (antibody-conjugated fluorophores, fluorescent dyes, and mRNA in-situ hybridisation), and analysis of non-specific light scatter.

Facility users have access to two 20+ parameter flow cytometric analysers, two cell sorters, an imaging cytometer – the Imagestream (please check equipment for details), and a dedicated analysis suit workstation complete with FlowJo, Cytobank and IDEAS software. After training users gain 24/7 access to analysers and can operate them on their own. Cell sorting is offered as a service – this powerful technique allows you to enrich a population of interest or source single cells of interest from cell suspension. Cells after this procedure are viable, re-suspended in media of your choice. Cells can be placed in different types of collection tubes or plates (24wp, 96wp, 384wp).

Please contact us if you would like to inquire about any of the services we offer or if you would like to arrange a visit to the lab.

 

Flow Cytometers (analysers)

Analysers are available for users 24/7. Please contact for details.

BD Fortessa X20 – has 5 lasers: 355 (UV), 405, 488, 561 and 640 nm. It has 18 fluorescent channels available. Cytometer is equipped with HTS (High Throughput System – 96wp reader).

Cytoflex LX – has 4 lasers 405, 488, 561, and 640 nm. It has 16 fluorescent channls available. Cytometer is equipped with a 96 well plate reader.

Cell Sorters

Sorts are performed by authorised operators only.

The flow lab is equipped with a jet-in-air hi-speed sorter (MoFlo Astrios from Beckman Coulter) and one cuvette cell sorter (Aria III from BD).

Astrios: equipped with 4 lasers: 405, 488, 561 and 640 nm and is able to detect up to 14 fluorescent parameters. This sorter is in a 2 class hood.

BD FACSAria III: equipped with 4 lasers: 405, 488, 561 and 635 nm and has 16 fluorescent channels.

We offer a range of nozzle options (70-200um) allowing high speed bulk sorts (up to 30 000 events per second), sorting of large cells, and plate sorting (single cell deposition) with index sort option.

 

Imaging Flow Cytometer

Amnis Imagestream MKII imaging flow cytometer –provides cutting edge high throughput microscopy to flow lab users after complimentary training. It is equipped with 4 lasers: 405, 488, 561 and 638 nm offering 9 fluorescent channels, dual brightfeild, and side scatter (cell complexity). The technology behind the machine allows users to take high resolution images (optical magnification of 20x, 40x or 60x) of every single event and combine flow data analysis with microscopy analysis simultaneously. Data can then be analysed on a dedicated analysis suite using the Amnis IDEAS software – training for this software is also provided.

Group members

  • Robert Hedley (Facility Manager)
  • Vasiliki Tsioligka

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