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

项目摘要 AD是一种复杂的、进行性的神经退行性疾病，并且是AD的最常见原因。 老年痴呆症它有一个明确的空间和时间进程;多行 有证据表明，在AD中看到的退化模式不是随机的，而是显示出一种 随着病理学的发展，几十年来，的机制 导致这些空间模式的原因是知之甚少， 微观分子机制与宏观解剖学变化的贡献 AD进展的驱动因素尚不清楚。 不幸的是，像MRI这样的全脑技术缺乏必要的分子对比， 解决这些问题。组织学可以揭示微观的，分子的 签名，但由于大比例尺的技术限制， 组织学，特别是3D组织学。这些限制使得很难绘制AD进展图 并限制了可能揭示AD关键线索的死后组织的有用性。 在本提案中，我们将通过提供按服务收费的分段、切片 安装，染色和成像平台，将提供非专业实验室的能力， 获得高质量的死后人脑三维连续切片重建， 半球可将样本发送至TissueVision，在那里将其按厚度进行切片 范围从50微米到2毫米，在切片之间交替进行块面成像， 便于3D重建。每个固定切片可用于进一步染色， 通过全载玻片成像（WSI）实现高分辨率数字化，并通过云服务于 客户端可选的3D建模。物理部分可以返回到客户端或存储 以便进行进一步的后续分析。 该提案结合了TissueVision在显微镜和3D组织学方面的专业知识， 类临床研究人员在印第安纳州大学和建立在现有的工作在TissueVision。的 切片捕获和幻灯片安装基于我们的 用于小鼠和大鼠大脑的TissueCyte 1600 FC STPT平台。我们还建立在一个强大的 与杰克逊实验室和MODEL-AD中心合作，使用现有的TissueVision Phase II NIA SBIR以绘制小鼠模型脑中的AD空间进展。我们将延长 与印第安纳州大学AD模型的临床合作者合作， 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，但拟议的技术将产生广泛的影响 从心脏生物学、传染病、癌症和基础医学， research.

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.