Correlative In Vivo Fluorescence and Micro-Computed Tomographic Imaging of Tissue Structure and Function

为了使人体和动物身体正常运作，组成它们的组织和器官必须组织得当（具有正确的组织结构）。这必须发生在许多尺寸尺度上，从单个细胞到大型器官，如肝脏，肾脏和骨骼，或供应器官的血管和神经。鉴于细胞和组织是动态的三维（3D）结构，我们需要了解它们如何在3D中组合在一起，以了解它们的结构，结构如何在组织内连接和变化。例如，在再生医学中，我们试图制造组织和器官来修复受损和患病的组织，并将身体恢复到原来的健康状态。细胞可以在其上生长并被引导的支架用于帮助将细胞组织成正确的结构。我们现在处于一个独特的位置，创造新的软组织和硬组织（如肝脏，神经组织，软骨，骨），以改善治疗，并更好地了解正常和异常的人类和动物的身体功能。同样，随着年龄的增长，我们身体的组成也会发生变化，能够跟踪和了解这种变化在活体动物中是如何发生的是很重要的，这样我们最终就可以掌握更好地治疗一系列疾病的知识，比如心脏病、肺病和癌症。我们正在申请一种成像系统，它将允许我们检查细胞，体内器官和组织（称为相关体内成像）以在连续时间点创建3D图像，该系统将允许我们标记和跟踪身体内的特定细胞和分子，使用高分辨率微计算机断层摄影（uCT）成像设备对感兴趣组织的堆叠进行成像。这使用X射线对大样本进行非破坏性成像，这种新仪器的设计使我们能够区分单个组织成分以及它们的一些特征如何随着时间的推移而变化，这是目前通过一种称为组织学的技术在二维（2D）中无法实现的，使用正常，该成像系统将增加南安普顿现有成像设施的范围，这些设施由12名专业成像人员提供支持，在生物医学成像单位（BIU）和u-VIS X射线成像中心（u-VIS）工作。现有的样品制备和生物图像解释方面的专业知识对于有效使用这些尖端成像技术至关重要。所有尺度的3D成像中的另一个问题是产生的非常大的数字图像数据集，每个数据集占用大量的存储空间，通常为50- 1000 Gb（每个相当于10-200个DVD！）。因此，关键是要有专门的工作人员，他们将运行并确保成像系统有效运行，并为所有用户提供可靠的简化服务。美国已同意资助一名研究支持工作人员，以支持和提供可持续的成像服务。提供组合的临床前相关体内成像系统，以及我们现有的先进计算和图像处理领导团队（BIU和u-VIS已经拥有最先进的计算硬件，软件和专业知识），将允许研究和了解组织结构并更好地了解组织功能。在这个项目中，我们将与我们在大学的合作者合作，进一步开发组织样本的相关体内成像应用领域，特别是包括3D数据集的处理和分析工作流程。我们在其原生3D背景下理解组织发育和形成的能力有可能在未来10-30年内改变人类健康。为了尽可能快速和安全地实现这一承诺，我们必须能够以纵向方式和组织的不同层次对体内生成的结构和组织进行成像。

For human and animal bodies to function properly, the tissues and organs that make them up must be organised properly (have the correct tissue architecture). This must happen across many size scales, from individual cells to large organs such as the liver, kidneys, and bones, or blood vessels and nerves supplying the organs. Given cells and tissues are dynamic three-dimensional (3D) structures, we need to see how they fit together in 3D to understand their architecture, how structures connect and vary within the tissue. For example, in Regenerative Medicine, we try to make tissues and organs to repair damaged and diseased tissues and restore the body to its original health. Scaffolds upon which the cells can grow and are guided are used to help organise the cells into the right structures. We are now in a unique position to create new soft and hard tissues (e.g. liver, neural tissue, cartilage, bone) to improve treatments and to better understand normal and abnormal human and animal body function. Similarly, in aging the composition of our bodies changes, and it is important to be able to track and understand how this happens in living animals, so that ultimately we can have the knowledge to better treat a range of diseases, like heart disease, lung disease and cancer.We are applying for an imaging system that will allow us to examine cells, organs and tissues within the body (called Correlative in vivo imaging) to create 3D images at successive time points, without having to kill the animal to look inside it. The system will allow us to label and follow specific cells and molecules within the body and to create 3D image stacks of tissues of interest with a high-resolution micro-computed tomographic (uCT) imaging device. This uses X-rays to non-destructively image large samples and the design of this new instrument allows us to distinguish individual tissue components and how some of their features change over time, in a way that is not currently possible in two dimensions (2D) through a technique called histology, using normal, light microscopy.The imaging system will add to the wide range of existing imaging facilities in Southampton that are supported by 12 expert imaging staff, employed at the Biomedical Imaging Unit (BIU) and the u-VIS X-ray Imaging Centre (u-VIS). The existing expertise in sample preparation and biological image interpretation is essential for these cutting edge imaging techniques to be used effectively. An additional problem in 3D imaging at all scales is the very large digital image datasets that are produced, each taking large amounts of storage, typically; 50-1000Gb (equivalent of 10-200 DVDs each!). On this account, it is pivotal to have dedicated staff in place who will run and make sure that the imaging system performs efficiently and can provide a reliable, streamlined service to all users. The UoS has agreed to fund a Research Support staff member to underpin and provide a sustainable Imaging Service. Provision of the combined preclinical correlative in vivo imaging system, together with our existing leadership team in advanced computing and image processing (state-of-the-art computing hardware, software and expertise already in place at BIU and u-VIS, will allow investigating and understanding tissue architecture and to understand better tissue function. In this project, we will work with our collaborators in the University to develop further correlative in vivo imaging application areas of tissue samples, specifically including processing and analysis workflows for the 3D datasets. Our ability to understand tissue development and formation in its native 3D context has the potential to transform human health over the next 10-30 years. In order to fulfil this promise, as quickly and safely as possible, it is essential we can image the generated structures and tissues in vivo in a longitudinal fashion and at different hierarchical levels of tissue organisation.