"Grid-Tape": A High-Throughput Platform for Brain Connectomics and Nanoscale Structural Analysis

“Grid-Tape”：用于脑连接组学和纳米级结构分析的高通量平台

• 批准号: 10078761
• 负责人: Ryan M Smith
• 金额: $ 54.02万
• 依托单位: LUXEL CORPORATION
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-07 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078761
• 关键词: Alzheimer' s Disease; Area; Automation; BRAIN initiative; Brain; Communities; Data; Data Analyses; Data Set; Development; Disease; Economics; Electron Microscope; Electron Microscopy; Excision; Film; Foundations; Funding; Geometry; Goals; Grant; Health; Human; Image; Imaging Techniques; Individual; Institutes; Laboratories; Lasers; Maps; Methods; Microscope; Modernization; Morphologic artifacts; Mus; Network-based; Neurodegenerative Disorders; Neurons; Neuropil; Neurosciences; Noise; Pathway interactions; Phase; Polymers; Preparation; Price; Production; Property; Protocols documentation; Research Personnel; Resolution; Role; Running; Sampling; Scanning; Schizophrenia; Science; Series; Signal Transduction; Speed; Stains; Standardization; Structure; Synapses; System; Technology; Thick; Thinness; Tissue Sample; Tissue Stains; Tissue imaging; Tissues; Transmission Electron Microscopy; United States National Institutes of Health; Visual Cortex; Work; autism spectrum disorder; base; brain tissue; cold temperature; cost; cost effective; data acquisition; data quality; field study; flexibility; improved; innovation; instrument; instrumentation; microscopic imaging; millimeter; nanoscale; nervous system disorder; petabyte; sample collection; sensor; tool; transmission process

The wiring diagram of brain circuits is one of the foundational and fundamental questions of modern neuroscience. Since the connectivity of these circuits critically influences their function, understanding the structure of these networks will have a major impact on understanding their role in health and disease. The primary challenge is extracting a circuit-sized volume at synaptic resolution, which requires large-scale electron microscopy imaging of thousands of sections of brain tissue. Until now, the slow speed of tissue sectioning and imaging has been a major limitation to the field of connectomics, requiring many years to acquire a dataset of this size. GridTape™, developed in Phase I, removes this bottleneck by allowing fast automated imaging of thousands of sections on a continuous tape inside a transmission electron microscope (TEM). This new reel- to-reel sample substrate leverages inherent speed and resolution advantages of camera-based TEM to provide higher data acquisition rates with lower cost than scanning electron microscope (SEM) alternatives. In Phase I, GridTape™ achieved a longstanding goal for connectomics with the successful acquisition of a cubic mm volume at 4 nm resolution, yielding a dataset of more than 2 petabytes. The data spanned 26,500 serial sections split between seven reels of GridTape™, and was imaged with five TEM microscopes running in parallel for six months at an effective imaging rate of 500 Mpixel/s. While the best multi-beam SEM alternative can achieve 3X the imaging rate of a single TEM with GridTape™, the cost is roughly 20X higher ($4-6M) and only a few multi-beam SEMs currently exist in the world; this drives the economics of volume-EM heavily in favor of an approach using multiple cheaper TEMs in parallel with GridTape™. With feasibility clearly demonstrated, Luxel will partner with Harvard in Phase II to improve a number of data quality issues for GridTape™. The technical challenges include reducing the background image noise from intrinsic structure in the support films, reducing costs for the thin film coating, mitigating image artifacts that arise from tissue cracks and folds, and expanding the film-covered slot areas to allow larger tissue samples without breakage. We plan to achieve these goals by developing alternative low-noise polymer films that also offer manufacturing scalability benefits including faster substrate removal. We will partner with connectomics researchers at Harvard and the Allen Institute to determine optimal tissue block preparation formulas and methods to maintain film tension that mitigate tissue cracks and folds.

大脑电路的接线图是现代的基础和基本问题之一 神经科学 网络的结构将对他们在健康和疾病中的作用产生重大影响 主要挑战是在突触分辨率下提取电路尺寸的体积，这需要大规模电子 到目前为止的显微镜成像。 成像一直是连接组领域的主要限制，要求许多Youys获取数据集 这种大小。 在透射电子显微镜（TEM）中连续胶带上的数千个部分。 to-reel样品基板利用基于相机TEM的固有速度和分辨率的优势来提供 比扫描电子显微镜（SEM）替代的较高的数据采集率。 GridTape™通过成功获得立方MM的连接组学实现了长期的连接目标。 4 nm分辨率的体积，产生超过2 pb的数据集。 截面分裂在Gridtape™的七个卷轴之间，并用五个TEM显微镜成像 平行六个月的有效成像速率为500 mpixel/s。 可以通过GridTape™实现单个TEM的成像率的3倍，成本大约高20倍（4-6m），并且成本 目前，世界上只有几个多光束SEM； 使用与Gridtape™并行的多个较便宜的TEM的方法。 卢克斯（Luxel GridTape™。 支撑膜，降低薄膜涂层的成本，减轻组织裂缝的图像图像伪影 折叠，扩大膜覆盖的插槽区域，以允许我们计划的较大的组织样品。 通过开发替代性低噪声聚合物膜来实现这些目标，这些薄膜也提供制造业 可扩展性好处，包括更快的基板。 哈佛大学和艾伦学院以确定最佳组织块制备公式的tomethods维持 减轻组织裂纹和褶皱的膜张力。