A Scalable Staining Strategy for Whole-Brain Connectomics.
A Scalable Staining Strategy for Whole-Brain Connectomics.
复制标题
全脑连接组学的可扩展染色策略。
DOI：
10.1101/2023.09.26.558265
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Lichtman,JeffW
中科院分区：
文献类型：
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作者：
Lu,Xiaotang;Wu,Yuelong;Schalek,RichardL;Meirovitch,Yaron;Berger,DanielR;Lichtman,JeffW
绘制哺乳动物大脑的完整突触连接将是变革性的，揭示感知，行为和记忆的基础途径。连续切片电子显微镜，通过使用四氧化锇膜染色，是理想的可视化细胞和突触连接，但在全脑样本，面临着与化学处理和体积变化的重大挑战。这些问题可能对超微结构质量和宏观组织完整性产生不利影响。通过利用延时X射线成像和大脑代理，我们开发了一种12步的协议，ODeCO，有效地渗透整个小鼠大脑，同时保留超微结构没有任何裂缝或碎片，这是构建第一个全面的小鼠大脑连接体的必要先决条件。
Mapping the complete synaptic connectivity of a mammalian brain would be transformative, revealing the pathways underlying perception, behavior, and memory. Serial section electron microscopy, via membrane staining using osmium tetroxide, is ideal for visualizing cells and synaptic connections but, in whole brain samples, faces significant challenges related to chemical treatment and volume changes. These issues can adversely affect both the ultrastructural quality and macroscopic tissue integrity. By leveraging time-lapse X-ray imaging and brain proxies, we have developed a 12-step protocol, ODeCO, that effectively infiltrates osmium throughout an entire mouse brain while preserving ultrastructure without any cracks or fragmentation, a necessary prerequisite for constructing the first comprehensive mouse brain connectome.