A Tool for Synapse-level Circuit Analysis of Human Cerebral Cortex Specimens.

人类大脑皮层样本突触级电路分析的工具。

• 批准号: 10670926
• 负责人: Jeff W Lichtman
• 金额: $ 59.45万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-13 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10670926
• 关键词: Animals; Bilateral; Biopsy; Biopsy Specimen; Brain; Brain Diseases; Brain imaging; Cell physiology; Cerebral cortex; Cognition Disorders; Collaborations; Collection; Computer software; Computers; Cortical Column; Custom; Data; Data Set; Deep Brain Stimulation; Development; Electron Microscope; Electron Microscopy; Electrons; Epilepsy; Epitopes; Excision; Goals; Human; Image; Immersion; Individual; Invertebrates; Label; Laboratories; Lead; Maps; Medical center; Mental disorders; Methods; Modification; Molecular; Molecular Profiling; Neurologic; Neurons; Neurosciences; Neurosurgeon; Neurosurgical Procedures; Obsessive-Compulsive Disorder; Operating Rooms; Operative Surgical Procedures; Organ; Osmium; Pathologic; Patients; Permeability; Phase; Preservation Technique; Privatization; Procedures; Protocols documentation; Psychiatric therapeutic procedure; Quality Control; Resolution; Rodent; Running; Sampling; Scanning; Scanning Electron Microscopy; Schizophrenia; Scientist; Site; Specimen; Speed; Stains; Standardization; Structure; Synapses; Techniques; Temporal Lobe; Thick; Tissue Sample; Tissues; Variant; Work; Writing; brain tissue; brain volume; cell type; connectome; density; developmental disease; excitatory neuron; frontal lobe; implantation; implementation tool; improved; inhibitory neuron; insight; microscopic imaging; millimeter; multiple datasets; nanobodies; nanoscale; nervous system disorder; neural circuit; neuropathology; neurosurgery; nonhuman primate; novel; parallelization; petabyte; preservation; programs; sample fixation; supercomputer; technique development; tool

Project Summary/Abstract The goal of this work is to facilitate synaptic level analysis of neurons and their interconnecting microcircuits in neurosurgical cerebral cortex biopsies from human patients. These full-thickness human cerebral cortex biopsies will be provided by neurosurgical colleagues from patients undergoing resective surgery or surgical implantation of leads for deep brain stimulation (DBS). Once we have demonstrated that the techniques and tools are sufficiently reliable, we will analyze neural circuits in samples from medical centers that study psychiatric and neurological disorders. In the initial phase we will: 1) optimize the removal of undamaged brain biopsies during neurosurgical procedures and transfer new techniques for immersion fixation and osmium staining to large (>5 cubic millimeter) fresh brain biopsies from human patients.; 2) we will optimize, with hardware and software changes, the speed and reliability of multibeam scanning electron microscopy image acquisition to automatically acquire synapse-level neural circuitry at petabyte scale in brain volumes that connect tens of thousands of neurons via hundreds of millions of synapses; 3) we will transfer methods to co-register molecular labels (for cell -types) with serial electron microscopy of the same human samples using very small novel immuno-probes that do not require permeabilization and hence, do not negatively impact the quality of the brain’s ultrastructure in order to identify ultrastructural correlates for each cell type; and 4) we will work with computer scientists at Argonne National Laboratory to develop a robust computational connectomics pipeline for stitching, alignment, segmentation, and storage of human brain circuits. This public platform will augment the efforts of a team at Google that is already begun working on our human samples. In the second phase, we will run many human biopsies through the image acquisition and analysis connectomic pipelines. We will use new software to compare circuit variability within and between individuals. We contend that detailed neural circuit analysis in human brain tissue that bridges scales from nanometers to millimeters is a prerequisite for understanding how the normal brain functions and discovering the pathological underpinnings of cognitive and developmental disorders. Our goal is that the methods we develop will be disseminated, becoming part of the toolbox for both neuropathology and fundamental human neuroscience.

项目摘要/摘要 这项工作的目标是促进神经元及其相互连接的微电路的突触水平的分析 人类病人的神经外科大脑皮层活检。这些全层的人类大脑皮层活检 将由接受切除手术或外科植入的患者的神经外科同事提供 用于脑深部刺激(DBS)的导联。一旦我们展示了这些技术和工具 足够可靠，我们将分析研究精神病学和医学中心的样本中的神经电路 神经紊乱。在初始阶段，我们将：1)优化未受损脑组织活检的移除 神经外科手术和转移浸泡固定和Os染色的新技术到大型(&gt；5 立方毫米)新鲜的人类患者脑活检；2)我们将通过硬件和软件进行优化 改变了多束扫描电子显微镜图像采集的速度和可靠性，实现了自动化 在连接数万人的大脑体积中获得PB级的突触级别神经电路 通过数以亿计的突触传递神经元；3)我们将转移方法来共同注册分子标记(针对细胞 -类型)，使用非常小的新型免疫探针对相同的人类样本进行连续电子显微镜检查， 不需要通透性，因此不会对大脑的超微结构质量产生负面影响 以确定每种细胞类型的超微结构相关性；以及4)我们将与计算机科学家合作， Argonne国家实验室将开发一种强大的计算连接管道，用于缝合、对齐、 人脑回路的分割和存储。这一公共平台将加强一个团队在 谷歌已经开始研究我们的人体样本。在第二阶段，我们将运行许多人类 活检通过图像采集和分析连接管道。我们将使用新的软件来比较 个体内部和个体之间的电路可变性。我们认为，人类大脑中详细的神经回路分析 连接从纳米到毫米尺度的组织是理解正常的 大脑功能和发现认知和发育障碍的病理基础。我们的 目标是我们开发的方法将被传播，成为神经病理学工具箱的一部分 和基础的人类神经科学。