Award details

Cutting-edge Spectral sorter to drive in-depth analysis of cell states and activity in complex tissue environments

Reference	BB/V019287/1
Principal Investigator / Supervisor	Dr Joanne Konkel
Co-Investigators / Co-Supervisors	Professor Stuart Allan, Professor Judith Allen, Professor David Bechtold, Professor David Brough, Professor Silvia Bulfone-Paus, Professor Kevin Couper, Professor Daniel Davis, Professor Kathryn Else, Professor Julie Gibbs, Dr John Grainger, Professor Richard Grencis, Dr Matthew Hepworth, Professor Tracy Hussell, Professor Karl Kadler, Dr Gloria Lopez-Castejon, Professor Andrew Loudon, Professor Andrew MacDonald, Dr Pawel Paszek, Professor Ian Roberts, Professor Andrew Sharrocks, Dr Joe Swift, Professor David Thornton, Professor Mark Travis
Institution	The University of Manchester
Department	School of Biological Sciences
Funding type	Research
Value (£)	481,495
Status	Completed
Type	Research Grant
Start date	01/08/2021
End date	31/07/2022
Duration	12 months

Abstract

Identifying different cell types at the single cell level is crucial to determine how they maintain tissue health. A vital technique in studying single cells in tissues is flow cytometry, which allows detection of specific cells by the markers they express. Populations of different cells can then be sorted based on fluorescent parameters and used in downstream applications such as RNA-sequencing, proteomics, and in vivo/vitro assays. Thus, flow cytometry allows new levels of biological understanding; it supports analysis of cells at unprecedented resolution, enhancing understanding of the immense cellular complexity of tissues and systems. As such, it is now necessary to divide cell populations into subsets using novel combinations of markers to allow for detailed dissection of cell biology. This represents a bottleneck for research at the University of Manchester; so many markers are required for cell identification that our current sorters cannot isolate the novel populations identified through higher-dimensional analysis. We currently have the ability to analyse up to 50 parameters in a sample. However, the current cell sorters available can isolate cell populations based on only 14 parameters. Advances in flow cytometry in the last decade struggled because of overlap between the emission spectra of fluorophores used to label antibodies. The newly released Bigfoot Cell Sorter from Propel Labs provides a paradigm shift by separating fluorescence signals from cells based on full spectral readout of fluorochromes. This enables increased flexibility and sensitivity compared to existing systems as spectral data acquisition allows expanded panels to be developed with currently available fluorophores. The Bigfoot spectral cell sorter would allow us to sort cell populations based on 45 parameters, an extensive advancement on our current sorting capacity. Thus, this sorter will substantially enhance our ability to study cellular function, plasticity and development.

Summary

Our body is composed of trillions of cells, which form the specialised tissues and organs that allow us to function normally. To understand how our body works, it is critical to understand the biology of our cells; how they communicate with each other, how they perform their specialised functions, and how they change as we age or are injured. Our tissues are made up of many different types of cell, so to study how specific types of cell work, it is vital that we can identify and isolate individual cells from within complex samples. A powerful technique to do this is flow cytometry, which allows researchers to identify different types of cells based on markers they express, and then subsequently isolate and purify these cells based on these markers. In this way researchers are able to isolate defined populations of cells for subsequent in-depth analysis of their function. This supports detailed insight into how cells develop and function to promote health, and how this changes during ageing or following injury. As such, isolation of cells using flow cytometric sorters underpins a plethora of research areas across biology aiding fundamental insight into how cells function to promote health. Isolation of specific and well-characterized populations of cells has become even more important in recent years; as there has been an explosion in our understanding of cell diversity. For most cell types examined, we now know that there are many different versions of what were initial thought to be the same cell. Thus, heterogeneity within a single population of cells has now been well described; what we now need to understand is how this cell diversity underpins and ensures health. This requires isolation of these different cell populations, which currently cannot be achieved at the University of Manchester. Here we request funds to purchase a Bigfoot Spectral Cell sorter, to allow isolation and purification of cells based upon 45-parameters. This will greatly facilitate theresearch of the applicant consortium speeding up progress of existing projects and making new projects possible. This equipment will support work from a variety of priority research areas including immunology, microbiome and microbiology research, the biology of aging, neuroscience, cell biology, developmental biology and systems biology. Immediately, acquisition of a Bigfoot sorter will benefit at least 24 groups of highly productive, and well regarded, scientists at the University of Manchester. This equipment will also enhance the training of the next generation of scientists, providing skills in cutting-edge spectral flow cytometry. In sum, the Bigfoot sorter will greatly progress the research of the consortium, enabling interdisciplinary research that will provide in-depth knowledge of the cellular mechanisms that safeguard health.

Committee	Not funded via Committee
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Advanced Life Sciences Research Technology Initiative (ALERT) [2013-2014]
Funding Scheme	X – not Funded via a specific Funding Scheme

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