Dendritic patch-clamp recording
Dendritic patch-clamp recording
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DOI：
10.1038/nprot.2006.164
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Haeusser, Michael
中科院分区：
文献类型：
--
作者：
Davie, Jenny T.;Kole, Maarten H. P.;Haeusser, Michael
膜片钳技术允许研究神经元的电兴奋性和离子通道的功能特性和密度。大多数神经元的膜片钳记录是从索马（单个神经元的最大结构）进行的，而形成大部分表面积并接收大部分突触输入的树突相对被忽视。该协议描述了在直接视觉控制下从脑切片中的神经元树突记录的技术。虽然基本的技术是类似的，用于体细胞修补，我们描述的细化和优化切片质量，显微镜光学，设置稳定性和电极的方法，需要最大限度地提高成功率为树突状记录。使用这种方法，可以实现膜片钳技术的所有配置（细胞贴附，由内而外，全细胞，由外而外和穿孔的补丁），即使是相对较远的树突，同时多电极树突记录也是可能的。该方案-从切片准备开始到第一次成功记录结束-可以在3小时内完成。
The patch-clamp technique allows investigation of the electrical excitability of neurons and the functional properties and densities of ion channels. Most patch-clamp recordings from neurons have been made from the soma, the largest structure of individual neurons, while their dendrites, which form the majority of the surface area and receive most of the synaptic input, have been relatively neglected. This protocol describes techniques for recording from the dendrites of neurons in brain slices under direct visual control. Although the basic technique is similar to that used for somatic patching, we describe refinements and optimizations of slice quality, microscope optics, setup stability and electrode approach that are required for maximizing the success rate for dendritic recordings. Using this approach, all configurations of the patch-clamp technique (cell-attached, inside-out, whole-cell, outside-out and perforated patch) can be achieved, even for relatively distal dendrites, and simultaneous multiple-electrode dendritic recordings are also possible. The protocol - from the beginning of slice preparation to the end of the first successful recording - can be completed in 3 h.