Abstract
Ion-selective microelectrodes can be used to report intracellular ion concentrations. The ion-selective barrels of microelectrodes are filled with a sensor cocktail containing several different components including an ion-selective molecule, sensor or exchanger, a solvent or plasticizer, lipophilic cation/anion additives, and a matrix to solidify the membrane. For many ions, the readymade membrane cocktail can be purchased, but the individual chemical components can be bought from suppliers and mixing the cocktail saves money. For commercially available liquid membrane cocktails the membrane matrix is often not included. For plants a matrix is essential for intracellular impalements because without it cell turgor will displace the liquid membrane from the electrode tip, giving decreased or even lost sensitivity. The matrix frequently used is a high molecular weight poly(vinyl chloride). This addition increases the electrical resistance of the electrode, slowing the response time of the electrode. The use of multi-barreled 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.
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References
Ammann D (1986) Ion-selective microelectrodes, principles, design and application. Springer, Berlin
Henriksen GH, Bloom AJ, Spanswick RM (1990) Measurement of net fluxes of ammonium and nitrate at the surface of barley roots using ion-selective microelectrodes. Plant Physiol 93:271–280
Miller AJ, Cookson SJ, Smith SJ, Wells DM (2001) The use of microelectrodes to investigate compartmentation and the transport of metabolized inorganic ions in plants. J Exp Bot 52:541–549
Delessert C, Wilson IW, Van der Straeten D, Dennis ES, Dolferus R (2004) Spatial and temporal analysis of the local response to wounding in Arabidopsis leaves. Plant Mol Biol 55:165–181
Miller AJ, Zhen R-G (1991) Measurement of intracellular nitrate concentrations in Chara using nitrate-selective microelectrodes. Planta 184:47–52
Walker DJ, Smith SJ, Miller AJ (1995) Simultaneous measurement of intracellular pH and K+ or NO −3 in barley root cells using triple-barreled ion-selective microelectrodes. Plant Physiol 108:743–751
Fry CH, Hall SK, Blatter LA, McGuigan JAS (1990) Analysis and presentation of intracellular measurements obtained with ion- selective microelectrodes. Exp Physiol 75:187–198
Sanders D, Slayman CL (1982) Control of intracellular pH. Predominant role of oxidative metabolism, not proton transport, in the eukaryotic microorganism Neurospora. J Gen Physiol 80:377–402
Acknowledgments
John Innes Centre is grant-aided by the Biotechnology and Biological Sciences Research Council (BBSRC) of the UK.
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Miller, A.J. (2013). Real Time Measurement of Cytoplasmic Ions with Ion-Selective Microelectrodes. In: Maathuis, F. (eds) Plant Mineral Nutrients. Methods in Molecular Biology, vol 953. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-152-3_16
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DOI: https://doi.org/10.1007/978-1-62703-152-3_16
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-151-6
Online ISBN: 978-1-62703-152-3
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