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用于哺乳动物中枢神经系统神经元膜片钳记录的薄片制备。

A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system.

作者信息

Edwards F A, Konnerth A, Sakmann B, Takahashi T

机构信息

Max-Planck-Institut für Medizinische Forschung, Abteilung Zellphysiologie, Heidelberg, Federal Republic of Germany.

出版信息

Pflugers Arch. 1989 Sep;414(5):600-12. doi: 10.1007/BF00580998.

Abstract

(1) A preparation is described which allows patch clamp recordings to be made on mammalian central nervous system (CNS) neurones in situ. (2) A vibrating tissue slicer was used to cut thin slices in which individual neurones could be identified visually. Localized cleaning of cell somata with physiological saline freed the cell membrane, allowing the formation of a high resistance seal between the membrane and the patch pipette. (3) The various configurations of the patch clamp technique were used to demonstrate recording of membrane potential, whole cell currents and single channel currents from neurones and isolated patches. (4) The patch clamp technique was used to record from neurones filled with fluorescent dyes. Staining was achieved by filling cells during recording or by previous retrograde labelling. (5) Thin slice cleaning and patch clamp techniques were shown to be applicable to the spinal cord and almost any brain region and to various species. These techniques are also applicable to animals of a wide variety of postnatal ages, from newborn to adult.

摘要

(1)本文描述了一种可对哺乳动物中枢神经系统(CNS)神经元进行原位膜片钳记录的制备方法。(2)使用振动组织切片机切出薄片,在其中可通过视觉识别单个神经元。用生理盐水对细胞胞体进行局部清洁,使细胞膜游离,从而在膜与膜片吸管之间形成高阻封接。(3)利用膜片钳技术的各种配置来证明可记录神经元和分离膜片的膜电位、全细胞电流和单通道电流。(4)膜片钳技术用于记录充满荧光染料的神经元。通过在记录过程中填充细胞或通过先前的逆行标记来实现染色。(5)薄片清洁和膜片钳技术被证明适用于脊髓和几乎任何脑区以及各种物种。这些技术也适用于从新生到成年的各种出生后年龄的动物。

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