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

技术的发展使得对细胞进行极其详细的研究，同时测量许多不同的分子（称为多参数分析），已经彻底改变了生物医学领域的研究。这些技术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有机体发育和生理，通过维持组织稳态、衰老或产生免疫反应。因此，在许多不同的研究领域，现在需要检查超过10种不同的分子（在这种情况下称为参数）来简单地识别感兴趣的细胞。例如，它可能需要研究多达15种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的单元的活动或状态。虽然这些多参数技术（包括流式细胞术、CyTOF和单细胞rna测序技术）非常强大，但它们的问题是，它们都需要将被检查的组织处理成单细胞悬浮液进行分析，并且丢失了有关组织中细胞来源的所有背景。我们对各种生物过程的了解越多，就越明显地发现，大多数生物事件（无论是组织稳态、免疫反应的产生、对损伤或疾病的反应）都是多因素的，涉及组织内不同细胞类型的相互作用。此外，细胞在组织/器官内的位置经常在影响应答结果方面起主要作用。因此，很明显，要真正了解特定细胞群如何促进生物过程，或生物过程在健康期间如何发展和调节，或在疾病期间如何修改，至关重要的是要补充诸如流式细胞术，细胞tof和单细胞转录组学等研究，并在组织内进行多参数调查（即研究生理组织结构内的细胞和生物过程）。从历史上看，这是不可能的，因为用于检查组织的传统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大约10倍的多参数组织学调查的能力，这在生物学研究中是可能的。在此申请中，我们请求资金购买Hyperion成像系统。Hyperion系统的购买将为我们的联盟（在案例支持中概述）以及曼彻斯特和英格兰西北部的其他研究人员提供一个步骤，以在生理组织环境中进行多参数成像调查。这将为在不同的BBSRC战略领域中整合的许多不同的生物过程提供基本的新见解，特别是在“理解生命规则”和“生物科学对健康的综合理解”主题中。

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.