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Electrochemical measurement of endogenously produced nitric oxide in brain slices using Nafion/o-phenylenediamine modified carbon fiber microelectrodes [An article from: Analytica Chimica Acta]
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PublisherElsevier
ISBN / ASINB000RR3IOQ
ISBN-13978B000RR3IO3
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This digital document is a journal article from Analytica Chimica Acta, published by Elsevier in 2005. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
Description:
The role of nitric oxide (?NO) as a regulatory diffusible molecule in the brain requires the evaluation of its concentration dynamics. In this work, we have developed microelectrodes suitable for real time electrochemical measurements of ?NO in vitro. Nafion and o-phenylenediamine were used to modify the surface of carbon fiber microelectrodes (8@mm diameter; ~100@mm tip length). Coating with Nafion was done at 170^oC and the o-phenylenediamine solution was electropolimerized on the carbon surface. ?NO peak potential (+0.78+/-0.03V versus Ag/AgCl) was determined by square wave voltammetry with ?NO solutions prepared from the-generating compound diethylenetriamine/nitric oxide (DETA/NO). Microelectrodes were calibrated by amperometry at a potential of +0.90V versus Ag/AgCl. They showed good sensitivity (954+/-217pA/@mM; n=6) and linearity to ?NO in the concentration range of 100-1000nM. They were also characterized in terms of detection limit (6+/-2nM, n=4), response time at 50% (1s), and selectivity against interferents, such as nitrite (780+/-84:1, n=6), ascorbic acid (750+/-187:1, n=6) or dopamine (18+/-2:1, n=6). Injections of 1mM l-glutamate, 1mM l-arginine, and 0.1mM N-methyl-d-aspartate did not produce changes in background current. Finally, the microelectrodes were used to measure ?NO concentration dynamics in rat hippocampal brain slices stimulated with l-glutamate and N-methyl-d-aspartate. Taken together, the data indicate that the microelectrodes exhibit the proper sensitivity and selectivity for studies of ?NO dynamics in brain slices (in vitro) and possibly in whole brain (in vivo) recordings.
Description:
The role of nitric oxide (?NO) as a regulatory diffusible molecule in the brain requires the evaluation of its concentration dynamics. In this work, we have developed microelectrodes suitable for real time electrochemical measurements of ?NO in vitro. Nafion and o-phenylenediamine were used to modify the surface of carbon fiber microelectrodes (8@mm diameter; ~100@mm tip length). Coating with Nafion was done at 170^oC and the o-phenylenediamine solution was electropolimerized on the carbon surface. ?NO peak potential (+0.78+/-0.03V versus Ag/AgCl) was determined by square wave voltammetry with ?NO solutions prepared from the-generating compound diethylenetriamine/nitric oxide (DETA/NO). Microelectrodes were calibrated by amperometry at a potential of +0.90V versus Ag/AgCl. They showed good sensitivity (954+/-217pA/@mM; n=6) and linearity to ?NO in the concentration range of 100-1000nM. They were also characterized in terms of detection limit (6+/-2nM, n=4), response time at 50% (1s), and selectivity against interferents, such as nitrite (780+/-84:1, n=6), ascorbic acid (750+/-187:1, n=6) or dopamine (18+/-2:1, n=6). Injections of 1mM l-glutamate, 1mM l-arginine, and 0.1mM N-methyl-d-aspartate did not produce changes in background current. Finally, the microelectrodes were used to measure ?NO concentration dynamics in rat hippocampal brain slices stimulated with l-glutamate and N-methyl-d-aspartate. Taken together, the data indicate that the microelectrodes exhibit the proper sensitivity and selectivity for studies of ?NO dynamics in brain slices (in vitro) and possibly in whole brain (in vivo) recordings.
