High-resolution whole-brain staining for electron microscopic circuit reconstruction
High-resolution whole-brain staining for electron microscopic circuit reconstruction
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DOI：
10.1038/nmeth.3361
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发表时间：
2015-06-01
期刊：
影响因子：
48
通讯作者：
Denk, Winfried
中科院分区：
文献类型：
--
作者：
Mikula, Shawn;Denk, Winfried
目前，只有电子显微镜能够提供完全重建神经元回路和单突触分辨率所需的分辨率。由于几乎所有行为都依赖于广泛分布在整个大脑中的神经计算，因此非常需要重建全脑回路，并最终重建整个大脑。然而，这些重建需要未分割的大脑为电子显微镜观察做好准备。在这里，我们描述了一种制剂，BROPA（通过连苯三酚介导的放大进行全脑还原锇染色），它可以以追踪神经元过程和识别它们之间的突触接触所需的分辨率保存和染色整个大脑的超微结构细节。使用串行块面电子显微镜 (SBEM)，我们测试了人类注释者长距离可靠地跟踪神经“线路”的能力以及检测突触接触的能力。我们的结果表明，BROPA 方法可以生产出适合重建整个小鼠大脑神经回路的制剂。
Currently only electron microscopy provides the resolution necessary to reconstruct neuronal circuits completely and with single-synapse resolution. Because almost all behaviors rely on neural computations widely distributed throughout the brain, a reconstruction of brain-wide circuits-and, ultimately, the entire brain-is highly desirable. However, these reconstructions require the undivided brain to be prepared for electron microscopic observation. Here we describe a preparation, BROPA (brain-wide reduced-osmium staining with pyrogallol-mediated amplification), that results in the preservation and staining of ultrastructural details throughout the brain at a resolution necessary for tracing neuronal processes and identifying synaptic contacts between them. Using serial block-face electron microscopy (SBEM), we tested human annotator ability to follow neural 'wires' reliably and over long distances as well as the ability to detect synaptic contacts. Our results suggest that the BROPA method can produce a preparation suitable for the reconstruction of neural circuits spanning an entire mouse brain.