Targeted axon-attached recording with fluorescent patch-clamp pipettes in brain slices
Targeted axon-attached recording with fluorescent patch-clamp pipettes in brain slices
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DOI：
10.1038/nprot.2012.061
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发表时间：
2012-06-01
期刊：
影响因子：
14.8
通讯作者：
Ikegaya, Yuji
中科院分区：
文献类型：
--
作者：
Sasaki, Takuya;Matsuki, Norio;Ikegaya, Yuji
了解中枢神经系统中轴突的生理学需要对完整轴突进行实验。该方案描述了如何在急性和培养的脑切片制备物中从窄轴突纤维（phi < 1 μ m）进行细胞附着记录（成功率接近50%）。通过使用荧光团涂覆的玻璃移液器和Nipkow圆盘共聚焦显微镜，荧光标记的轴突可以在在线光学控制下视觉上靶向。在细胞附着的配置，轴突动作电位细胞外记录为单位样，尖锐的负电流。轴突形态标记和细胞贴附记录可在1-2 h内完成。记录至少稳定30 min。
Understanding the physiology of axons in the central nervous system requires experimental access to intact axons. This protocol describes how to perform cell-attached recordings from narrow axon fibers (phi < 1 mu m) in acute and cultured brain slice preparations (with a success rate of similar to 50%). By using fluorophore-coated glass pipettes and Nipkow disk confocal microscopy, fluorescently labeled axons can be visually targeted under online optical control. In the cell-attached configuration, axonal action potentials are extracellularly recorded as unit-like, sharp negative currents. The axon morphology labeling and cell-attached recordings of axons can be completed within 1-2 h. The recordings are stable for at least 30 min.