AN APPARATUS FOR RECORDING SYNAPTIC POTENTIALS FROM NEURONAL CULTURES USING VOLTAGE-SENSITIVE FLUORESCENT DYES
AN APPARATUS FOR RECORDING SYNAPTIC POTENTIALS FROM NEURONAL CULTURES USING VOLTAGE-SENSITIVE FLUORESCENT DYES
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DOI：
10.1016/0165-0270(91)90159-w
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发表时间：
1991-07-01
影响因子：
3
通讯作者：
PINE, J
中科院分区：
文献类型：
--
作者：
CHIEN, CB;PINE, J
电压敏感染料提供了无创多细胞电活动记录的前景，因此应该有助于研究培养神经元小网络的突触相互作用。我们设计并建立了一个用于记录来自大鼠颈上神经节(SCG)的1-15个神经元的显微培养系统，使用的是苯乙烯类的电压敏感荧光染料。该装置包括一个标准的倒置荧光显微镜；一个带有光反馈调节器的汞弧灯；一个带有独立光电二极管探测器和极低噪声放大器的256像素光纤摄像机；以及一个基于个人计算机的数据采集系统。它的暗噪声和光照波动足够低，以至于在这些细胞的典型荧光水平下，它受到散粒噪声(检测的固有物理极限)的限制。从SCG神经元记录下来，信噪比高到足以在不进行信号平均的情况下看到大的阈值下突触电位。这台仪器对于研究脊椎动物神经元培养中的长期突触可塑性是有用的，它的几个特征应该适用于其他制备的光学记录。
Voltage-sensitive dyes offer the promise of noninvasive multicell recording of electrical activity, and should therefore be useful for studying the synaptic interactions of small networks of cultured neurons. We have designed and built a system for recording from microcultures of 1-15 neurons from the rat superior cervical ganglion (SCG), using voltage-sensitive fluorescent dyes of the styryl class. The apparatus comprises a standard inverted epifluorescence microscope; a mercury arc lamp with an optical feedback regulator; a 256-pixel fiber-optic camera with individual photodiode detectors and very low-noise amplifiers; and a personal computer-based data acquisition system. Its dark noise and illumination fluctuations are low enough that at typical florescence levels for these cells, it is limited by shot noise (the inherent physical limit of detection). Recording from SCG neurons, the signal-to-noise ratio is high enough to see large subthreshold synaptic potentials without signal averaging. This apparatus should be useful for studying long-term synaptic plasticity in cultures of vertebrate neurons, and several of its features should apply to optical recording from other preparations.