Long-Term Optical Access to an Estimated One Million Neurons in the Live Mouse Cortex.
Long-Term Optical Access to an Estimated One Million Neurons in the Live Mouse Cortex.
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DOI：
10.1016/j.celrep.2016.12.004
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发表时间：
2016-12-20
期刊：
影响因子：
8.8
通讯作者：
Schnitzer MJ
中科院分区：
文献类型：
--
作者：
Kim TH;Zhang Y;Lecoq J;Jung JC;Li J;Zeng H;Niell CM;Schnitzer MJ
神经科学的一个主要技术目标是能够在活体大脑中对单个细胞进行询问。通过创建背侧颅骨的弯曲玻璃替代物和用于其安装的手术方法，我们开发了一种慢性小鼠制备物，其提供了对新皮层背侧表面上的估计800，000 - 1，100，000个个体神经元的光学访问。术后组织学研究显示，与对照小鼠中的胶质细胞活化相当。在表现出在皮质锥体神经元中表达Ca 2+指示剂的小鼠中，我们使用落射荧光宏观镜在新皮质上进行Ca 2+成像，并估计每只小鼠在多个焦平面上可接近25，000 - 50，000个单独的神经元。双光子显微镜显示了整个新皮层的树突状形态，允许对单个细胞进行延时成像，并通过连续平铺产生了每只小鼠> 100万个可接近神经元的估计值。这种方法支持各种光学技术，并能够研究行为小鼠中超过30个新皮层区域的细胞。Kim等人提出了一种长期成像的准备方法，其中弯曲的玻璃窗取代了小鼠的背侧颅骨。这种方法能够在行为小鼠的新皮层上对神经元动力学进行大规模的Ca 2+成像，并通过双光子显微镜产生估计>1百万个光学可接近的神经元。
A major technological goal in neuroscience is to enable the interrogation of individual cells across the live brain. By creating a curved glass replacement to the dorsal cranium and surgical methods for its installation, we developed a chronic mouse preparation providing optical access to an estimated 800,000–1,100,000 individual neurons across the dorsal surface of neocortex. Post-surgical histological studies revealed comparable glial activation as in control mice. In behaving mice expressing a Ca2+ indicator in cortical pyramidal neurons, we performed Ca2+ imaging across neocortex using an epi-fluorescence macroscope and estimated that 25,000–50,000 individual neurons were accessible per mouse across multiple focal planes. Two-photon microscopy revealed dendritic morphologies throughout neocortex, allowed time-lapse imaging of individual cells, and yielded estimates of >1 million accessible neurons per mouse by serial tiling. This approach supports a variety of optical techniques and enables studies of cells across >30 neocortical areas in behaving mice. Kim et al. present a preparation for long-term imaging in which a curved glass window replaces the mouse dorsal cranium. This method enables large-scale Ca2+ imaging of neuronal dynamics across neocortex in behaving mice and yields an estimated >1 million optically accessible neurons by two-photon microscopy.