An Automated platform for 3D reconstruction and profiling of post-mortem Alzheimer's Disease brains

项目总结 AD是一种复杂的进行性神经退行性疾病，是导致 老年人的痴呆症。它有一个清晰的时空进程；多条线 有证据表明，阿尔茨海默病的退变模式不是随机的，而是表现出一种 随着几十年来病理学的发展，具有特征性的解剖序列。其作用机制 导致这些空间格局的原因是人们对其知之甚少，以及关于 微观分子机制与宏观解剖学变化的关系 推动AD进展的因素仍不清楚。 不幸的是，像MRI这样的全脑技术缺乏必要的分子对比度和 回答这些问题的决心。组织学可以揭示微观的、分子的 签名，但由于大比例的技术限制，仅限于较小的2D剖面 组织学，特别是3D组织学。这些限制使绘制AD进程图变得困难 并限制了可能揭示有关AD的关键线索的尸检组织的有用性。 在本提案中，我们将通过提供按服务收费的分区Slice来解决这一障碍 安装、染色和成像平台将为非专业实验室提供 获得高质量的死后人脑连续切片三维重建 半个半球。可以将标本送到TIseVision，在那里它们将按厚度进行切片 从50微米到2毫米不等，切片之间交替进行块面部成像以帮助 促进3D重建。每个安装的部分都可用于进一步染色和 使用全幻灯片成像(WSI)以高分辨率进行数字化，并通过云服务到 具有可选3D建模功能的客户端。物理部分可以返回给客户端或存储 以作进一步的后续分析。 这项建议将TIssueVision的显微镜和3D组织学专业知识与World- 印第安纳大学一流的临床研究人员，并建立在TIssueVision现有工作的基础上。这个 截面捕获和幻灯片安装基于我们的 TIseCyte 1600FC STPT平台适用于小鼠和大鼠的大脑。我们还建立在强大的 与Jackson Labs和Model-AD中心以及现有TIseVision阶段的协作 II NIA SBIR绘制小鼠模型脑中AD的空间进展图。我们将延长 与印第安纳大学Model-AD的临床合作者合作，他们将 提供死后人体组织，并提供组织学指导和反馈 质量。虽然我们在这个项目中的重点是AD，但提议的技术将产生广泛的影响 涉及范围广泛的学科，从心脏生物学、传染病、癌症和基础 研究。

PROJECT SUMMARY AD is a complex, progressive neurodegenerative disease and is the most common cause of dementia among the elderly. It has a clear spatial and temporal progression; multiple lines of evidence show that the degeneration patterns seen in AD are not random but show a characteristic anatomical sequence as the pathology advances over decades. The mechanisms that cause these spatial patterns are poorly understood and basic questions about the relative contributions of microscopic molecular mechanisms versus macroscopic anatomical changes in driving AD progression remain unclear. Unfortunately, whole brain techniques like MRI lacks the necessary molecular contrast and resolution to answer these questions. Histology can reveal the microscopic, molecular signatures, but is constrained to small 2D sections due to technical limitations with large scale histology, particularly 3D histology. These limitations make it difficult to map AD progression and limit the usefulness of post-mortem tissue that might reveal crucial clues about AD. In this proposal we will address this barrier by providing a fee-for-service sectioning, slice mounting, staining, and imaging platform that will offer non-specialized labs the ability to obtain high quality 3D serial section reconstructions of post-mortem human brains up to entire hemispheres. Specimens can be sent to TissueVision where they will be sectioned at thicknesses ranging from 50 microns to 2 mm, with alternating block face imaging between sections to help facilitate 3D reconstruction. Each mounted section is available for further staining and digitization at high resolution with Whole Slide imaging (WSI) and served over the cloud to the client with optional 3D modeling. The physical sections can be returned to the client or stored for further follow-up analyses. This proposal combines expertise in microscopy and 3D histology at TissueVision with world- class clinical researchers at Indiana University and builds on existing work at TissueVision. The section capture and slide mounting is based on existing technology contained within our TissueCyte 1600FC STPT platform for mouse and rat brains. We also build on a strong collaboration with Jackson Labs and the MODEL-AD center with an existing TissueVision Phase II NIA SBIR to map AD spatial progression in mouse model brains. We will extend the collaboration to clinical collaborators at the Model-AD at Indiana University who will be supplying post-mortem human tissue and providing guidance and feedback on the histological quality. While our focus is on AD in this project, the proposed technology will have broad impact across a wide range of disciplines, from cardiac biology, infectious diseases, cancer, and basic research.