文献类型：期刊文献

中文题名：双管离子选择性微电极制备方法研究

英文题名：Preparation of double-barreled ion-selective microelectrodes

作者：李进海[1];薛琳[2];周桥[1];黄岚[1];王忠义[1]

第一作者：李进海

机构：[1]中国农业大学信息与电气工程学院,北京100083;[2]北京联合大学智慧城市学院,北京100101

第一机构：中国农业大学信息与电气工程学院,北京100083

年份：2021

卷号：37

期号：16

起止页码：24-30

中文期刊名：农业工程学报

外文期刊名：Transactions of the Chinese Society of Agricultural Engineering

收录：CSTPCD;;EI(收录号：20214711217794);Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金项目(61571443)。

语种：中文

中文关键词：微电极;试验;双管离子选择性微电极;硅烷化;制备方法

外文关键词：microelectrodes;test;double-barreled ion-selective microelectrodes;silanization;preparation method

摘要：双管离子选择性微电极被广泛应用于植物细胞外离子流速和细胞内离子活度的测量,但双管离子选择性微电极(Ion-Selective Microelectrode,ISME)的制备过程繁琐,不可控因素多,制备成功率低。针对存在的问题,该研究提出了一种简易、快速的双管ISME制备方法。首先,介绍了双管微电极制备、硅烷化和电极尖端灌充液态离子交换剂(Liquid Ion Exchanger,LIX)的具体流程;其次,对制备的双管ISME的能斯特斜率和响应时间进行了测试。试验结果表明,使用蒸汽硅烷法对双管微电极进行硅烷化处理,最优硅烷化温度、二甲基二氯硅烷剂量和硅烷化时间分别为150℃、45μL和90 min;制备的双管氢离子、钾离子、钙离子、氯离子选择性微电极的能斯特斜率分别为54.08、56.51、27.08和-58.80 mV/dec;4种双管ISME的响应时间介于0.20~0.42 s之间。研究结果表明,由该研究制备方法制作的双管ISME,可以满足植物细胞外离子流速和细胞内离子活度信息检测的要求。双管ISME的快速制备,降低了离子选择性微电极技术的应用难度,将有利于植物电生理检测试验的进行和离子选择性微电极技术在农作物育种、生理抗逆、植物营养吸收与同化等研究领域的应用。
Ion-Selective Microelectrode(ISME) technology has widely been used to evaluate the plant electrical activity in mesoscopic space-time scale, including dynamic measurements on extracellular ion fluxes and intracellular ion activities. But the interference of electromigration or membrane potential changes has posed a great challenge on the traditional single-barreled ISME during the electrophysiological experiments, particularly on the accuracy of measured data. In this study,a double-barreled ISME monitoring scheme was proposed to eliminate the influence of potential drift on the measurement for more accurate information of ion flux. A simple and rapid preparation process of double-barreled ISMEs fabrication was also developed to widen the application of ion-selective electrode technology. Septum Theta(a borosilicate glass capillary with two cavities) was used to fabricate the improved double-barreled microelectrode during preparation, particularly on the silanization and liquid ion exchanger filling in the microelectrode tip of double-barreled ISME. The experimental measurements were conducted for the Nernst slope and response time of double-barreled ISMEs. A standardized feasibility plan was provided for the preparation and performance testing of double-barreled ISMEs. Experimental results showed that the optimal silanization temperature, dimethyldichlorosilane dosage, and silanization time were 150 ℃, 45μL, and 90 min, respectively, when the double-barreled microelectrodes were silanized by the steam silane. In this case, the double-barreled ISME that filled 100 μm liquid ion exchanger in the tip was reciprocated in the test solution at a frequency of 0.5 Hz for 30 min, where the remaining amount of liquid ion exchanger was observed under the microscope. The remaining amount of liquid ion exchanger was(100±0) μm in the double-barreled potassium ion, calcium ion, and hydrogen ISMEs’ tip. In double-barreled chlorine ISMEs,the remaining amount of liquid ion exchanger in the tip was(90±8.2) μm. The Nernst slopes of double-barreled hydrogen ion,potassium ion, calcium ion, and chlorine ISMEs were 54.08, 56.51, 27.08, and-58.80 mV/dec, respectively. The measured Nernst slope of ISMEs was more than 90% of the theoretical value suitable for the requirements of the application. The response time of ion-selective electrodes with different liquid ion exchangers was between 0.2 and 0.42 s. Therefore, 0.5 Hz was a reasonable vibration frequency for the measurement of extracellular ion fluxes with the double-barreled ISMEs. The experimental results demonstrated that the double-barreled ISME developed by the improved preparation can well meet the requirements of ISME technology, thereby effectively capturing the extracellular ion fluxes or intracellular ion activities of plant cells. Consequently, the facile preparation of double-barreled ISME can be expected to greatly reduce the experimental difficulty of ISME fabrication. The finding can also provide a great contribution to acquire experimental data of plant physiological detection and applications in agricultural engineering, crop breeding, physiological stress tolerance, and cellular nutrition.