Establishment of a cutting-edge imaging modality to enable multi-parameter analyses within tissues

The development of technologies that allow the study of cells in extreme detail, measuring many different molecules simultaneously (called multi-parameter analysis), has revolutionised research within the biomedical field. These technologies have demonstrated the complexity and extreme cellular heterogeneity (variation) that exists within and underpins biological processes, spanning tissue and organismal development and physiology, through to maintenance of tissue homeostasis, aging or generation of an immune response. As a consequence, in many different research areas it now requires examination of more than 10 different molecules (in this context referred to as parameters) to simply identify a cell of interest. For example, it can require studying up to 15 different parameters to identify different immune cell subsets. Moreover, it can require 5-10 parameters to accurately define the activity or status of a cell of interest.

Whilst these multi-parameter technologies (which include techniques called flow cytometry, CyTOF and single cell RNA-sequencing), have been extremely powerful, the problem with them is that they all require that the tissue under examination is processed into single cell suspensions for analysis, and all context regarding where in the tissue the cell came from is lost. The more we reveal regarding various biological processes, the more obvious it becomes that most biological events (whether tissue homeostasis, generation of an immune response, response to injury or disease), are multi-factorial, involving the interaction of different cell types within the tissue. Moreover, the location of the cells within the tissue / organ frequently plays a major role in influencing the outcome of the response. Consequently, it has become clear that to truly understand how a particular cell population contributes to a biological process, or how a biological process develops and is regulated during health, or how it is modified during disease, it is critical to complement investigations such as flow cytometry, CyTOF and single cell transcriptomics, with multi-parameter investigations within the tissue (i.e. studying cells and biological processes within the physiological tissue structure). Historically this has not been possible as traditional imaging approaches for examining tissues have only allowed the study of up to 5 different molecules at a time, which means researchers have not even been able to identify their cell of interest within the tissue environment, let alone investigate how the cell interacts with other cells or define where it is located within the tissue structure.

To support multi parameter biological examinations within a physiological tissue environment, Fluidigm has recently developed the Hyperion Imaging mass cytometer system that allows the concurrent imaging of up to 37 parameters within an individual tissue section. Thus, the Hyperion system fundamentally changes, by approximately 10 fold, the power of multi-parameter histological investigations that are possible within biological research. In this application we request funds to purchase a Hyperion imaging system. The purchase of the Hyperion system will provide a step change in the ability of our consortium (outlined in the case for support), as well as other researchers at Manchester, and within the North West of England, to perform multi-parameter imaging investigations within physiological tissue environments. This will provide fundamental new insights into many different biological processes integral within different BBSRC strategy areas, particularly within the "understanding the rules of life" and the "bioscience for an integrated understanding of health" themes.

Technologies that enable high-dimensional cellular phenotyping, such as flow cytometry, CyTOF and single cell transcriptomics underpin research within many areas of Biomedical science, including those within key BBSRC strategic themes. However, an inherent limitation of these methodologies is that they provide no physiological information on the spatial context of identified cells within tissues or the interrelationship of cells with the tissue microenvironment. Thus, although they have revealed the complexity and heterogeneity evident within biology, these methodologies do not resolve the roles of cells (or molecules) during biological processes. Consequently, one of the critical next steps in biomedical research must be the integration of high dimensional cellular phenotyping investigations, such as flow cytometry and CyTOF, with imaging modalities that provide complementary, multi-parameter and quantitative physiological tissue-context.

Researchers at Manchester currently have no capacity to perform high dimensional quantitative analysis of tissues (the current capacity is 5 parameter immunofluorescence staining). Thus, the current histological and bioimaging capabilities at Manchester are not compatible with many modern research programmes that require deep profiling of tissues to firstly identify complex cellular phenotypes (often requiring >10 parameters), and then to resolve the compartmentalisation and spatial relationship between cells of interest and the tissue environment. As such, in this application we request funds to purchase the Fluidigm Hyperion system, which is a transformative new system for immunohistochemical analysis that enables the multiplex imaging of up to 37 parameters. Purchase of the Hyperion will thus provide a step change in the capacity of scientists at Manchester to study and understand, with necessary multi-parameter and spatial context, various different biological processes within BBSRC priority research areas.

This application requests a Hyperion Imaging CyTOF machine that will support the research of a large consortium of users at the University of Manchester and which will be of benefit to the wider research community in the North West of England. The results obtained using the Hyperion, studying myriad biological processes spanning different BBSRC priority areas, will be of significant interest to researchers in academia and industrial sectors as well as clinicians working with patients with relevant conditions. Through the various engagement and outreach activities, the research using the Hyperion will also have impact with the general public, school and university students.

The purchase of the Hyperion will immediately benefit researchers in the consortium. The Hyperion will allow the researchers to revolutionise the nature of the investigations undertaken within ongoing research programmes, which will dramatically enhance the impact of their research. This will have a direct and immediate benefit in terms of publications and will underpin planned grant applications. The Hyperion will also facilitate and promote new interdisciplinary research within the academic research community at University of Manchester, and more broadly in the North West of England and elsewhere. Results and data analysis tools that will be developed, particularly in the area of quantitative biology, to exploit the capabilities of the Hyperion system, will be shared for use of all researchers.

As the Hyperion represents a transformative technology that fundamentally alters the nature of quantitative and multi-parameter investigations possible in cells within their physiological tissue environment, the purchase of the machine will have significant impact for biotechnology. The purchase of the machine will catalyse development and utilisation of new biotechnology tools to multiplex the study of intracellular RNA, proteins and structures with high sensitivity, which will lead to generation of resources that will be useful for the wider scientific community. We expect a high potential impact in the biotechnology area and will actively search for relevant systems/companies to share our knowledge

The results to be obtained using the Hyperion system will provide fundamental new insights into the biology of complex cellular, tissue / organ and organismal processes. We anticipate that the results will have impact for identifying new mechanisms involved in health and disease, which will be of interest to researchers in translational science, clinicians, and those scientists in the industrial sectors.

We will communicate our results through various public engagement activities and outreach events. Images generated from the Hyperion will be colourful, intuitive, attractive and make science more accessible. Our results will be used to educate members of the public how research within physiological tissue environments provides critical insight into a range of different biological processes. With reference to the projects outlined in the case for support, this will help raise awareness of the science underlying the impact of aging, how tissue homeostasis is controlled, the role and importance of commensal microbes in the body, the biology of the brain, and how and why the body is controlled though a circadian cycle. However, in the longer term as the breadth of research performed using the Hyperion increases, we expect our results to have long term impact in public engagement across many areas of Biomedical science.