SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction
SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction
复制标题
DOI：
10.1038/nn.3447
复制
发表时间：
2013-08-01
影响因子：
25
通讯作者：
Imai, Takeshi
中科院分区：
文献类型：
--
作者：
Ke, Meng-Tsen;Fujimoto, Satoshi;Imai, Takeshi
我们报告了一种水基光学清除剂 SeeDB，它可以在几天内清除固定的大脑样本，而不会淬灭多种类型的荧光染料，包括荧光蛋白和亲脂性神经元示踪剂。我们的方法在清除过程中保持恒定的样品体积，这是保持细胞形态完整的重要因素，并促进神经元回路的定量重建。结合双光子显微镜和优化的物镜，我们能够对小鼠大脑从背侧到腹侧进行成像。我们使用 SeeDB 描述了与小鼠嗅球中共同肾小球相关的姐妹二尖瓣细胞的近乎完整的接线图。我们发现姐妹二尖瓣细胞之间树突布线模式的多样性，我们的结果为这些神经元的非冗余气味编码提供了解剖学基础。我们简单而有效的方法可用于对成人和发育中大脑的完整形态结构进行大规模成像。
We report a water-based optical clearing agent, SeeDB, which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. Our method maintained a constant sample volume during the clearing procedure, an important factor for keeping cellular morphology intact, and facilitated the quantitative reconstruction of neuronal circuits. Combined with two-photon microscopy and an optimized objective lens, we were able to image the mouse brain from the dorsal to the ventral side. We used SeeDB to describe the near-complete wiring diagram of sister mitral cells associated with a common glomerulus in the mouse olfactory bulb. We found the diversity of dendrite wiring patterns among sister mitral cells, and our results provide an anatomical basis for non-redundant odor coding by these neurons. Our simple and efficient method is useful for imaging intact morphological architecture at large scales in both the adult and developing brains.