Multi-Parameter Flow Cytometry for Deep Molecular Phenotyping of Cell State Across the Life Course

Multi-parameter flow cytometry and cell and nuclei sorting are indispensable in modern biological sciences, enabling advanced single-cell level analysis of biological, environmental, and clinical samples and allowing deep molecular phenotyping and isolation of selected cells for further in-depth investigations bringing sophisticated understanding of how a single cell or microbe, a tissue or a whole organism work and interact with their environment. A better understanding of these mechanisms will guide us to more effective solutions to global health and sustainability challenges.

This multiuser proposal builds on a pressing need for a system capable of spectral high-performance, multi-parameter fluorescence-activated cell/nucleus sorting that will serve a cross-disciplinary consortium of research groups in life sciences at the University of Southampton and external academic and industrial collaborators within the South of England, investigating the mechanisms underpinning human health and aging, and food and environmental sustainability. The system will integrate significant advancements in cell and nuclear sorting, allowing the highest power, performance, and flexibility, enabling researchers across diverse disciplines in biological and computational sciences and bioengineering, with the highest relevance to those interested in temporal and spatial changes in cell states within tissues and microbial and environmental systems and how this defines their heterogeneities, functional relationships, and interaction with their environment.

The scientific interests of the flow cytometry consortium span areas of biology, ranging from healthy ageing, cell and developmental biology, bioengineering, sustainable agriculture, biofilms, antimicrobial resistance, host-microbe interaction, and environmental sustainability. This includes research into microbial communities and biofilms, for applications, from combatting antimicrobial resistance to food security.

The instrument would enable deep molecular phenotyping of heterogeneous bacteria within a biofilm to decipher the mechanisms underpinning their interactions and the basis of their resistance at a single-cell resolution, with a view of finding approaches to better disperse them. Other researchers aim to discover factors that influence early mammalian development and tissue growth throughout life combining three-dimensional organotypic cultures (referred to as organoids) with single cell/nucleus sequencing and artificial intelligence; and uncover the molecular and cellular organisation of the nervous system that underlies our ability to learn and remember.

Researchers would be able to accurately isolate heterogeneous single breast stem cells and characterise the molecular mechanisms that dictate how they proliferate and contribute to breast development and architecture. The equipment would allow researchers to distinguish between heterogeneous immune cells to enhance our understanding of their contribution to inflammation, in health and autoimmune diseases.

It would enable researchers to accurately isolate skeletal cells from the sea urchin and sea cucumbers and their relatives to discover the evolutionary mechanisms of biomineralization. This is also relevant to researchers exploring cellular mechanisms of growth and cell fate in osteoarthritis using nuclei extracted from calcified tissues including growth plate cartilage.

The advanced multi-parameter sorter will act as a hub that catalyses new cross-disciplinary collaborations between its diverse user base, from basic biology to bioengineering and biotechnology. This will be facilitated through established regional partnerships with the BBSRC National Biofilm and Innovation Centre (NBIC) and South Coast Biosciences Doctoral Partnership (SoCoBio DTP).