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

项目概要/摘要 这项工作的目标是促进神经元及其互连微电路的突触水平分析 来自人类患者的神经外科大脑皮层活检。这些全层人类大脑皮层活检 将由接受切除手术或手术植入的患者的神经外科同事提供 用于深部脑刺激（DBS）的引线。一旦我们证明了技术和工具是 如果足够可靠，我们将分析来自研究精神科和精神科的医疗中心样本中的神经回路 神经系统疾病。在初始阶段，我们将：1）优化期间未损坏脑活检的去除 神经外科手术并将浸没固定和锇染色新技术转移到大（> 5 立方毫米）来自人类患者的新鲜脑活检。； 2）我们将通过硬件和软件进行优化 变化，多束扫描电子显微镜图像采集的速度和可靠性自动 获得脑容量中 PB 级的突触级神经回路，连接数以万计的神经元 神经元通过数亿个突触； 3）我们将转移方法来共同注册分子标签（对于细胞 -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 不需要透化，因此不会对大脑超微结构的质量产生负面影响 以确定每种细胞类型的超微结构相关性； 4）我们将与计算机科学家合作 阿贡国家实验室将开发强大的计算连接组学管道，用于缝合、对齐、 人脑回路的分割和存储。这个公共平台将增强团队的努力 谷歌已经开始研究我们的人类样本。在第二阶段，我们将运行许多人力 通过图像采集和分析连接组管道进行活检。我们将使用新软件进行比较 个体内部和个体之间的回路变异性。我们认为，对人脑的详细神经回路分析 桥接纳米尺度到毫米尺度的组织是理解正常组织如何运作的先决条件 大脑功能并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 和基础人类神经科学。

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.