A platform for brain-wide imaging and reconstruction of individual neurons.
A platform for brain-wide imaging and reconstruction of individual neurons.
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DOI：
10.7554/elife.10566
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发表时间：
2016-01-20
期刊：
影响因子：
7.7
通讯作者：
Chandrashekar J
中科院分区：
文献类型：
--
作者：
Economo MN;Clack NG;Lavis LD;Gerfen CR;Svoboda K;Myers EW;Chandrashekar J
轴突乔木的结构控制着来自单个神经元的信号如何在哺乳动物大脑内传递。然而，很少有远程投射神经元的树干被完整重建，因为直径小至 100 nm 的轴突在分散在许多毫米组织的目标区域中形成树干。我们引入了一个用于完整组织体积的高分辨率三维荧光成像的平台，该平台能够实现长距离轴突轴的可视化和重建。该平台依赖于集成了组织振动切片机的高速双光子显微镜和一套用于大规模图像数据的计算工具。我们通过重建单个小鼠大脑运动皮层中多个神经元的轴突轴来展示这种方法的威力。 DOI：http://dx.doi.org/10.7554/eLife.10566.001 神经细胞或神经元长距离相互传输电脉冲。这些信号通过称为轴突的高度分支的神经纤维传播，轴突比人的头发细约一百倍，并且可以延伸到整个大脑。从头到尾追踪神经元的轴突可以帮助解释单个神经元和大脑区域如何长距离传递信号。小鼠大脑包含大约 7000 万个神经元，追踪大脑内许多神经元的轴突是一个具有挑战性的问题。解决这个问题需要一种以足够高的细节对整个大脑进行成像的方法，以明确地解析和跟踪整个大脑中单个神经元的轴突。伊科诺莫，克拉克等人。现在展示了一种对与整个小鼠大脑一样大的组织样本进行三维成像的方法。该系统是完全自动化的，其工作原理是首先对样本暴露表面附近的一层组织进行成像，然后切下与已成像体积相对应的组织切片。然后重复这些步骤，直到整个样本都成像；对于整个小鼠大脑来说，这大约需要一周的时间，并产生大约 30 TB 的图像。 Economo、Clack 等人的进展可以揭示神经元如何以前所未有的精确度进行长距离通信。该方法现在可用于生成神经元及其长距离连接的综合数据库。这样的数据库将有助于模拟大脑中神经回路的作用，并为研究特定行为期间大脑活动的实验设计提供信息。 DOI：http://dx.doi.org/10.7554/eLife.10566.002
The structure of axonal arbors controls how signals from individual neurons are routed within the mammalian brain. However, the arbors of very few long-range projection neurons have been reconstructed in their entirety, as axons with diameters as small as 100 nm arborize in target regions dispersed over many millimeters of tissue. We introduce a platform for high-resolution, three-dimensional fluorescence imaging of complete tissue volumes that enables the visualization and reconstruction of long-range axonal arbors. This platform relies on a high-speed two-photon microscope integrated with a tissue vibratome and a suite of computational tools for large-scale image data. We demonstrate the power of this approach by reconstructing the axonal arbors of multiple neurons in the motor cortex across a single mouse brain. DOI: http://dx.doi.org/10.7554/eLife.10566.001 Nerve cells or neurons transmit electrical impulses to each other over long distances. These signals travel through highly branching nerve fibers called axons, which are about one hundred times thinner than a human hair, and can extend across the entire brain. Tracing the axon of a neuron from start to end can help to explain how individual neurons and brain areas communicate signals over long distances. A mouse brain contains approximately 70 million neurons, and tracing the axons of many neurons within a brain is a challenging problem. Tackling this problem requires a method for imaging entire brains in high enough detail to unambiguously resolve and follow axons from individual neurons across the brain. Economo, Clack et al. now demonstrate such a method for three-dimensional imaging of tissue samples as large as the whole mouse brain. This system is fully automated and works by first imaging a layer of tissue near the exposed surface of a sample, and then cutting off a slice of tissue that corresponds to the volume that has been imaged. These steps then repeat until the entire sample has been imaged; this takes about a week for a whole mouse brain and produces about 30 terabytes of images. Economo, Clack et al.’s advance can uncover how neurons communicate over long distances with an unprecedented level of precision. The method can now be used to generate a comprehensive database of neurons and their long distance connections. Such a database would aid efforts to model the roles of neural circuits in the brain, and inform the design of experiments to study brain activity during particular behaviors. DOI: http://dx.doi.org/10.7554/eLife.10566.002