Flow Facility Equipment

BD FACSDiscover™ S8 Cell Sorter

Derive greater insights and unlock new discoveries using image-enabled sorting and spectral flow cytometry.

The instrument has 5 lasers (UV: 349nm - 30mW, Violet: 405nm - 50mW, Blue: 488nm - 100mW, Yellow/Green: 561nm - 50mW, Red: 637nm - 100mW) and 78 fluorescent APD detectors. It also comes with three imaging detectors for blue laser excitable fluorochromes. BD CellView™ Image Technology images cells with the electronic and optical components used in flow cytometers. This unique technology makes it possible to produce images without a camera, enabling real-time imaging for analysis and sorting.

The BD FACSDiscover™ comes with three different nozzle sizes (85, 100 and 130nm diameter) and can sort into 1.5mL microtubes, 2.0mL and 5.0mL tubes as well as 6-, 24-, 48-, 96- and 384- well plates.

Index sorting for both spectral and imaging parameters is available.

The Cell Sorter is equipped with a fully integrated custom-tailored Class II hood making it suitable for CL2 work.

The S8 in our facility is BD’s 100^th FACS Discover™ S8.

Cytek Aurora Cell Sorter

The Aurora Cell Sorter allows users to transfer their panels directly from the Aurora analysers and sort rare populations.

The Aurora Cell Sorter takes cell sorting to the next level. The instrument comes with 5 lasers (UV: 355nm, 20mW, Violet: 405nm, 100mW, Blue: 488nm, 50mW, Yellow/Green: 561nm, 50mW, Red: 640nm, 80mW) which allow the detection of 3 scattering parameters and 64 fluorescent channels. Like the Aurora system, the Aurora CS provides the benefits of Full Spectrum Profiling. The system delivers high resolution helping to distinguish challenging cell populations, such as cells with high autofluorescence or dimly expressed biomarkers. The sorter comes with a variety of nozzles (70, 100, 130um) and allows up to 6-way sorting into 1.5 mL tubes, up to 4 ways into 5 mL tubes and sorting into 96-well plates. Index sorting is also available. It sits in a custom-made Class II hood allowing sorting of CL2 samples.

This is the first Aurora CS to be installed in the UK

Thermo Fisher Bigfoot x 3

The Bigfoot is a spectral cell sorter. It can carry out complex, high parameter analysis and sorting.

The Bigfoot has 6-9 lasers (340nm, 405nm, 445nm, 488nm, 532nm, 561nm, 594nm, 640nm and 785nm) and 55 fluorescence parameters. It is capable of carrying out spectral and conventional flow cytometry. The Bigfoot can bulk sort at comparable speeds to other jet-in-air sorters (for example the BD Influx) and can sort up to 6 populations simultaneously.

Plate sorting on the Bigfoot is incredibly quick due to the capability to sort multiple streams into a plate. A 96-well plate can be sorted in a few seconds. Other capabilities of the machine include straight-down sorting into plates, bulk sorting into multiple types of collection tubes at the same time, and sorting into 10x plates.

BD FACS Aria Fusion

5 lasers (488nm, 405nm, 561nm, 640nm, 355nm, 18 colours. The Fusion is in a Class 2 hood which allows sorting of human cell lines and human primary cells/tissues. This machine, funded by the BBSRC, can be used for plate sorting and index sorting

BD FACS Jazz x 3

3 lasers (488nm, 561nm, 640nm), 6 colours. The Jazz sorters are high-speed jet-in-air sorters that are ideal for sorting large numbers of cells, for example for CRISPR screening. They are also well-suited to sorting of smaller panels for example for fluorescent protein experiments. The Jazz sorters are capable of sorting 2 populations simultaneously. The machines can also do plate sorting and index sorting. The one of our Jazz sorters is housed in a biosafety cabinet for containment level 2 work.

Cytek Aurora Spectral Analyser x 3

The Cytek Aurora measures the emitted fluorescence of dyes across the whole spectra using a range of detectors. The signal is stitched together to create a full spectrum signature for each fluorophore. Gathering data from all detectors allows the Aurora to resolve dyes that conventional cytometers cannot, so allowing a greater number of fluorochromes to be used in combination.

The Aurora has 5 lasers (UV: 355 nm, 20 mW, Violet: 405 nm, 100 mW, Blue: 488 nm, 50 mW, Yellow/Green: 561 nm, 50 mW, Red: 640 nm, 80 mW), 3 scatter channels and 64 fluorescence channels. This allows complex, high parameter analysis to be carried out.

Flat-top beam shaping allows for high flow rates without compromising on data resolution and there is also a module for analysis of samples from 96- well plates. The instruments have an ASL for auto sampling and a small particle detector

Sony ID7000™

This is the first 7 laser ID7000 in the UK bringing unique flow cytometric capabilities to users of the facility.

The Sony ID7000™ Spectral cell analyser is a high parameter spectral cytometer. The system has 7 spatially separated lasers (320nm, 355nm, 405nm, 488nm, 561nm, 637nm, 808nm) and 186 detectors (FSC/SSC+184 Fluorescent parameters). It uses spectral technology that allows the use of more fluorophores including combinations not possible on conventional machines, such as FITC and GFP.

It is easy to use thanks to a comprehensive software suite with automated workflows and comes with an autosampler with agitation and cooling to enable walkaway operation. The samples can be loaded on 96-well plates (standard height flat/V/U bottom, half deep V bottom, deep U bottom), 384-well standard flat bottom plates, and 5mL (12x75mm) polystyrene tubes.

ThermoFisher Attune CytPix

The Attune CytPix allows the user to analyse their populations combining flow cytometry with brightfield imaging.

The Attune CytPix flow cytometer combines acoustic focusing with a high-speed brightfield camera, allowing you to measure morphology and flow cytometry parameters. The instrument has four lasers (Violet: 405nm, 100mW, Blue: 488nm, 100mW, Yellow: 561nm, 100mW, Red: 637nm, 140mW) and can detect up to 16 parameters including Forward and Side Scatter.

The acoustic focusing allows the user to run up to 1mL of sample volume in one minute (up to 65,000 events/sec) without any loss in resolution. It also comes with a fully automated plate loader.

BD LSR Fortessa5 x 2

These instruments have five spatially separated laser (355nm, 405nm, 488nm, 561nm and 640nm) which allow 18 colours to be simultaneously measured as well as 2 scatter parameters. They are equipped with a 561nm laser, which allows detection of red fluorescent proteins and better excitation of PE fluorochromes. Utilising this laser line also reduces spillover and compensation with other channels such as FITC. It also has a UV laser, allowing scientists to use the new BD Brilliant UV dyes.

The Fortessas are also equipped with a High-throughput sampler, which can run 96 and 384 well plates. Since purchase these instruments have proved very popular and are our most regularly used analysers.

Bio-Rad ZE5 Cell Analyser

Bio-Rad’s ZE5 Cell analyser allows users to take their analysis to another dimension. Not only is it a 5 laser (355nm, 405nm, 488nm, 561nm and 640nm) 30 parameter machine but it can be used for high-throughput analysis using tubes or plates. This machine allows simultaneous detection of 27 fluorescent and 3 scatter parameters (FSC and SSC from 488nm laser plus FSC from 405nm laser).

The filter configuration enables users to utilise a range of commercially available fluorochromes, allowing multi-colour phenotyping above the Fortessa’s capabilities whilst also giving the user more flexibility of dye combinations. The machine can be used in ‘stat’ mode to run individual tubes, but can easily be used with racks of 5ml and Eppendorf tubes to allow ease of use and walk away analysis.

Analysis of 96 well plates can be done in around 15 minutes making it perfect not only for in depth phenotyping studies but also screening. There is also a 384 well plate capability.

Cytek Amnis Imagestream MkII

4 laser (488nm, 405nm, 561nm, 640nm). This imaging cytometer produces multiple high-resolution images of every cell directly in flow, giving quantitative information about each cell in up to 12 channels simultaneously.

The Imagestream is an image cytometer that allows quantitative data to be acquired as well as images of every event passing through the instrument to be recorded. This machine can analyse up to 5000 cells/sec. This clever machine allows the user to look at features of cells such as co-localisation, staining location, cell-cell location, spot counting and cell death analysis.

Members of the core facility are available to help set up and optimise assays and help with post-acquisition analysis of the data.