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使用多管微电极测量细胞内离子浓度。

Measuring intracellular ion concentrations with multi-barrelled microelectrodes.

作者信息

Miller Anthony J, Smith Susan

机构信息

Department of Disease and Stress Biology, John Innes Centre, Norwich, UK.

出版信息

Methods Mol Biol. 2012;913:67-77. doi: 10.1007/978-1-61779-986-0_4.

Abstract

Ion-selective microelectrodes can be used to measure intracellular ion concentrations. The use of multi-barrelled electrodes enables the identification of the cellular compartment. For example, the inclusion of a pH-selective electrode enables the cytoplasm and vacuole to be distinguished. The ion-selective barrels of microelectrodes are filled with a sensor cocktail containing several different components. An ion-selective molecule, sensor or exchanger. Membrane solvent or plasticizer. Additives, e.g., lipophilic cation/anion. Membrane matrix to solidify the ion-selective membrane; essential for measurements in plant cells with a cell wall and turgor. For many ions, the ready-made membrane cocktail can be purchased, but the individual chemical components can be bought from suppliers and mixing the cocktail oneself is cheaper. For commercially available liquid membrane cocktails, the membrane matrix is not normally included. A matrix is needed if the microelectrodes are to be used in plants because cell turgor will displace a liquid membrane from the electrode tip, thereby changing or eliminating the sensitivity to the measuring ion. The matrix used is usually a high molecular weight poly(vinyl chloride), but can include other polymers, such as nitrocellulose for additional strength.

摘要

离子选择性微电极可用于测量细胞内离子浓度。使用多管电极能够识别细胞区室。例如,包含pH选择性电极可区分细胞质和液泡。微电极的离子选择性管内填充有一种传感器混合液,其中包含几种不同成分。一种离子选择性分子、传感器或交换剂。膜溶剂或增塑剂。添加剂,例如亲脂性阳离子/阴离子。膜基质以使离子选择性膜固化;这对于在具有细胞壁和膨压的植物细胞中进行测量至关重要。对于许多离子,可以购买现成的膜混合液,但各个化学成分可从供应商处购得,自行混合混合液更便宜。对于市售的液膜混合液,通常不包含膜基质。如果要在植物中使用微电极,则需要基质,因为细胞膨压会将液膜从电极尖端挤出,从而改变或消除对测量离子的灵敏度。所使用的基质通常是高分子量聚氯乙烯,但也可包括其他聚合物,如用于增加强度的硝化纤维素。

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