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

通过同时测量许多不同的分子(称为多参数分析)，能够对细胞进行极其详细的研究的技术的发展，使生物医学领域的研究发生了革命性的变化。这些技术已经证明了生物过程中存在的复杂性和极端的细胞异质性(变异)，这些生物过程跨越组织和生物发育和生理，一直到维持组织动态平衡、衰老或产生免疫反应。因此，在许多不同的研究领域，现在需要检查10多个不同的分子(在本文中称为参数)才能简单地识别感兴趣的细胞。例如，它可能需要研究多达15个不同的参数来识别不同的免疫细胞亚群。此外，它可能需要5-10个参数来准确定义感兴趣单元的活动或状态。虽然这些多参数技术(包括流式细胞术、细胞飞行时间和单细胞RNA测序技术)非常强大，但它们的问题是，它们都要求将被检查的组织处理成单细胞悬液进行分析，并且丢失了所有关于细胞来自哪里的上下文。我们对各种生物过程的揭示越多，就越明显地发现，大多数生物事件(无论是组织内稳态、免疫反应的产生、对损伤或疾病的反应)都是多因素的，涉及组织内不同类型细胞的相互作用。此外，细胞在组织/器官中的位置经常在影响反应结果方面发挥重要作用。因此，很明显，为了真正了解特定细胞群体如何促进生物过程，或生物过程如何在健康期间发展和调节，或在疾病期间如何修改，至关重要的是用组织内的多参数调查(即研究生理组织结构内的细胞和生物过程)来补充诸如流式细胞术、细胞飞行时间和单细胞转录组等研究。从历史上看，这是不可能的，因为传统的检查组织的成像方法一次只能研究多达5个不同的分子，这意味着研究人员甚至无法在组织环境中识别他们感兴趣的细胞，更不用说研究细胞如何与其他细胞相互作用或确定它在组织结构中的位置。为了支持生理组织环境中的多参数生物检查，Fluidigm最近开发了Hyperion成像质量细胞仪系统，该系统允许在单个组织切片内同时成像多达37个参数。因此，Hyperion系统从根本上改变了生物学研究中可能进行的多参数组织学研究的力量，大约增加了10倍。在此应用程序中，我们申请购买Hyperion成像系统的资金。购买Hyperion系统将使我们的财团(在支持案例中概述)以及曼彻斯特和英格兰西北部的其他研究人员在生理组织环境中执行多参数成像研究的能力发生重大变化。这将为BBSRC不同战略领域，特别是“理解生命规则”和“综合理解健康的生物科学”主题中的许多不同生物过程提供根本性的新见解。

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.