A rapid and non leaky way for preparation of the sharp intracellular recording microelectrodes

Masoud Fereidoni , Yaghoub Fathollahi , Mahyar Janahmadi , Iran Godarzi
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Abstract

To fill microelectrodes using backfilling method needs excessive time approximately 4–6  h. It is often difficult to fill microelectrodes without damage or leakage. A main problem is bubble formation in microelectrodes which has an impact on the electrical properties of the electrode and thus it influences the quality of the recording. Based on Archimede's principle there is a force within a solution which pushes insoluble material with a lower specific gravity upward and outside of the solution. Centrifugation can increase the force to eliminate the bubbles.

We designed a microelectrode holder to protect microelectrode sensitive tips from mechanical damage due to the gravity tensions; it can help to eliminate the bubbles easily and simultaneously in 10  min or less.

The tests were performed for 2000, 4000, and 8000  rpm centrifugation each one for 3, 6 and 12  min duration respectively, it was found that the bubbles were completely eliminated at 8000  rpm for 6–12  min and there were no significant differences for resistance, and the number of leaky or damaged electrodes between the two methods.

In the new design of devices, the materials used and the design of the holder are simple and the approach is applicable to many laboratories worldwide.

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一种快速无漏制备细胞内记录微电极的方法
采用回填法填充微电极需要超长的时间,大约需要4 ~ 6小时,通常很难填充微电极而不损坏或不泄漏。微电极中的气泡形成是一个主要问题,它会影响电极的电学性能,从而影响记录的质量。根据阿基米德原理,溶液中存在一种力,它将比重较低的不溶物质向上推至溶液外。离心可以增加力来消除气泡。我们设计了一种微电极支架,以保护微电极敏感尖端免受重力张力造成的机械损伤;它可以在10分钟或更短的时间内轻松消除气泡。分别进行了2000、4000、8000 rpm离心3、6、12 min的实验,结果发现,在8000 rpm离心6 - 12 min时,气泡被完全消除,两种方法在电阻、漏电极和损坏电极的数量上没有显著差异。在新装置的设计中,所用的材料和支架的设计都很简单,该方法适用于全球许多实验室。
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