Advancing the state-of-the-art using spectral flow cytometry

Lead Research Organisation: University of Birmingham
Department Name: Institute of Immunology & Immunotherapy


The spectral cytometer will advance several research programs at the University of Birmingham beyond the state-of-the-art, including how the immune system responds to diverse challenges such as infection, cancer, vaccines, and autoimmunity (Infections and Immunity), and blending fundamental basic science with clinically relevant approaches to develop translational research programs (Translational Research). Many of our research groups address global health problems, from an expanding ageing population to life-threatening infections that hinder health and progress in Southern Africa (Global Health). Across all our research themes, they are united in attempting to develop more stringent and powerful methodology to address ongoing research problems. There are multiple challenges that the spectral cytometer will help us overcome, from reducing assay variability to removing confounding factors such as autofluorescence (Better methods, better research). The spectral cytometer will also allow access to the state-of-the-art technology for our MRC Advanced-interdisciplinary models (AIM) PhD programme. A spectral cytometer will therefore help us advance our research beyond state-of-the-art in a number of MRC priority areas.

Our current cytometers are the workhorse machines of the facility with each running at full capacity. One of the major issues our cytometry department faces is significant and rising user demand. A major reason for this is that the current technology can only practically incorporate up to 18 markers, meaning samples must be split to run all parameters. This has a detrimental impact on the productivity of our researchers who may have to wait for booking space to become available, and the heavy use places strain on our machines. A spectral flow cytometer will significantly alleviate current pressures in the facility. This is not only achieved by providing a new cytometer but provides the option of running more complex samples (e.g., greater panel size) in a shorter time frame. In addition to alleviating current pressures on our existing equipment, the spectral cytometer will enable our institute to develop new research strengths in our MRC-aligned research, as well as support complementary research in cancer immunology, inflammatory arthritis, and host-pathogen interaction (funded via CRUK, Wellcome Trust, Kennedy Trust and BBSRC).

The lead applicant Prof Kai Toellner has served as academic lead for the TechHub flow cytometry unit for 5 years and has considerable expertise in flow cytometry and analysis. Kai Toellner is a world expert in B cell and germinal centre immunology. The investigator team comprises a strong and diverse group of Co-I's including early to mid-career researchers (Drummond, Bending, Long, Croft) and experienced MRC programme-level funded researchers (Moss, Anderson). These groups have a track record of delivering high quality and impactful science across many areas of the MRC remit.

Technical Summary

Conventional flow cytometry utilises filters with set bandwidths that distinguish the peak emissions of common fluorophores. The need for filters sets a physical upper limit on the number of fluorochromes that can be used; the invention of spectral flow cytometry, which captures the entire emission spectrum of each fluorochrome before undergoing a process of spectral unmixing, circumvents this limitation. Our researchers face the challenge of identifying rare cell populations from low cellularity (e.g., murine tumours) or precious samples (e.g., patient biopsies). Often, those samples are highly auto-fluorescent and difficult to analyse by conventional cytometry. This application sets out the case for the purchase of a spectral flow cytometer for use by researchers at the University of Birmingham and across the Midlands. A spectral flow cytometer will increase our cell identification resolution for each sample (from 18 to 50 simultaneous parameters), leading to a significant acceleration in discoveries while improving data quality. Importantly, acquisition of this new equipment will require minimal new reagents since the latest spectral cytometers are compatible with current reagents meaning this equipment will provide excellent value for money for users. While users for this machine will primarily be MRC-funded researchers based at the University of Birmingham, access will opened to all academics from the Midlands as part of the UoBFCF commitment to the Midlands Innovation programme.


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