Large-volume en-bloc staining for electron microscopy-based connectomics.
Large-volume en-bloc staining for electron microscopy-based connectomics.
复制标题
用于基于电子显微镜的连接组学的大批量En-Bloc染色。
DOI：
10.1038/ncomms8923
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发表时间：
2015-08-03
影响因子：
16.6
通讯作者：
Helmstaedter M
中科院分区：
文献类型：
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作者：
Hua Y;Laserstein P;Helmstaedter M
大规模的连接组学需要对神经元组织块进行致密染色以进行电子显微镜检查（EM）。在这里，我们报告了一个大体积的密集整块EM染色协议，克服了染色梯度，这到目前为止基本上限制了三维（3D）EM的重建体积。我们的方案提供了来自小鼠新皮质的直径至少为1 mm的致密可重建组织块。通过放松对精确的地形样本定位的限制，它使神经元回路的相关功能和结构分析变得现实。 神经元回路（或连接组学）的大规模致密重建需要用于大体积致密整体电子显微镜（EM）染色的方法。在这里，作者开发了一种用于对直径至少为1 mm的小鼠新皮层组织块进行染色的方案，从而能够进行相关的功能和结构电路分析。
Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic. Large-scale dense reconstruction of neuronal circuits (or connectomics) requires methods for large-volume dense en-bloc electron microscopy (EM) staining. Here the authors develop a protocol for staining tissue blocks from mouse neocortex sized at least 1 mm in diameter, enabling correlated functional and structural circuit analyses.