Electrochemical and UV-Vis Spectroscopic Measurements of Nitric Oxide in Fibroblasts and Astrocytes

✍ Scribed by Mariusz Macherzynski; Wiktor Bilecki; Jerzy Gorecki; Ryszard Przewlocki; Janusz Golas

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 122 KB
• Volume: 12
• Category: Article
• ISSN: 1040-0397
• DOI: 10.1002/1521-4109(200009)12:13<1046::aid-elan1046>3.0.co;2-7

✦ Synopsis

Two complementary analytical methods were applied to measure nitric oxide production in two cell cultures of ®broblasts and astrocytes: UV-vis spectroscopic assay of nitrites based on Griess reaction and direct, electrochemical determination of nitric oxide with use of a 5 mm diameter disk porphyrinic sensor. The nitrite measurement was used in order to get total quantitative information on NO production in a speci®ed time. Application of an electrochemical sensor allowed observation of the dynamics and stabilization of nitric oxide production in real time, in the vicinity of a culture of stimulated cells. In experiments, cell suspension was placed just above the sensor surface or cells were grown directly on the modi®ed electrode. Current curves were recorded after base current stabilization and injection of L-arginine the last remaining cosubstrate in the system. Collected data enabled one to estimate the local concentration of nitric oxide produced in the culture of ®broblasts to be 70 nM.