Spectral Flow Cytometry for the Midlands

Flow cytometry is a method used in the biological sciences to count and profile mixtures of cells in a liquid. For example, immunologists use flow cytometry to work out the different types of immune cells in a blood sample. They can use flow cytometry to identify the different cell types, count them and profile them. Antibodies labelled with fluorescent dyes can be used to identify cells with specific features.

The technology underlying flow cytometry is continually improving. In the 1980s, equipment utilised a single laser and analysis of 2 colours (i.e. cell features). Over the years, the number of lasers and the possible colour combinations has increased with the conventional flow cytometers now offering 5+ lasers and analysis of up to 20 colours. Spectral flow cytometry, by changing the underlying technology from detection of a specific wavelength range for a particular fluorophore to measurement of the full optical spectrum for each particle, is now pushing the boundaries of the technology even further. The Cytek Aurora, requested in this application, is a newly released five-laser spectral flow cytometer that can measure up to 40 colours. State-of-the-art optics and software enables this overall high performance. Together the improvements offered by this spectral flow cytometer mean that researchers can measure more features within a single sample, leading to improved characterisation of cell populations. Unlike other high performance flow cytometers, this spectral flow cytometer can be applied to the study of a broad range of samples, including plant cells, bacteria, yeast, small particles and mammalian cells.

The new equipment will be located in the University of Nottingham's Flow Cytometry facility and will extend the capability of that facility. The facility, managed by Dr David Onion, has an excellent track record in delivering cutting-edge flow cytometry facilities to researchers across the UoN and beyond. It has established mechanisms for access, charging and an extensive client list both at the University of Nottingham, across the Midlands Innovation group of universities, and in industry that have indicated keen interest in gaining access to this technology. The facility has well-established processes in place for managing access and data, developing costing models for charging users and training new users.

The Cytek Aurora, requested in this application, is a newly released five-laser spectral flow cytometer (including a UV laser - 355nm; Violet laser - 407nm; Blue laser - 488nm; Yellow laser - 561nm; and a Red laser - 640nm) with 64 channels of detection. The inclusion of state-of-the-art optics and software enables overall high performance. Together these features will enable measurement of up to 40 parameters simultaneously, the ability to resolve highly overlapping stains and application in the broadest range of sample types from small extracellular vesicles, yeast and bacteria to plant and mammalian cells. The capability to acquire large high parameter datasets at high throughput is also transformational.

Examples of research that will benefit from access to multi-dimensional data generated by this spectral flow cytometer include studies with emerging antibody panels including 30+ antibodies for more comprehensive immunological analysis of samples from humans or animals, studies with panels of fluorescent stains - including overlapping stains - that aim to document phenotypic heterogeneity in yeasts in response to stress, and studies requiring resolution of small bacteria in order to analyse numerous chromosomal and synthetic reporter gene driven tags in combination with antibody and aptamer staining for multi-parameter biomarker and bio-product monitoring within large populations of cells.

The new equipment will be located in the University of Nottingham's Flow Cytometry facility. Dr Lucy Fairclough, the facility's academic lead, and Dr David Onion, the facility's technical manager, will use well-established processes already in place within the facility to manage access to the new equipment across the University. Their existing association with the Midlands Innovation Flow Cytometry group (founded by Dr Fairclough in 2015), will be used to extend use of the spectral flow cytometer and to offer training to the widest possible user base.

Acquisition of a spectral flow cytometer at the University of Nottingham will have wider impact across a variety of stakeholder groups:

- Researchers: Access to spectral flow cytometry will ensure that our facility remains at the forefront of this specialism in the UK and through international research collaborations. Formal training in high parameter, high throughput spectral flow cytometry will also have positive impacts on researchers. The University of Nottingham Flow Cytometry Facility offers a training to students (in particular BBSRC-funded DTP students), early career researchers, academic and industrial colleagues across the Midlands. These efforts have recently been boosted through funding from Midlands Innovation to enable new and experienced scientists to develop cutting-edge skills in flow cytometry and keep the Midlands at the fore-front of scientific research.

- The University of Nottingham: local access to a spectral flow cytometer will facilitate research in priority areas of the institution's research strategy, including Green Chemicals and Future Foods.

- The Midlands: researchers across the Midlands are interested in gaining access to this equipment, as demonstrated in the Letters of Support attached to this application. The integral involvement of the applicant team from the University of Nottingham in managing the Midlands Innovation Flow Cytometry group will help to ensure promotion and use by the widest possible group.

- The general public will also benefit in the longer term through impacts of the research in a wide range of industries, which are then able to design new and improved products and processes.