Measurement of nitric oxide in single cells and tissue using a porphyrinic microsensor.
This unit describes the preparation and applications of porphyrinic sensors for quantitative measurement of nitric oxide (NO) in single cells and in tissues. The determination of NO is based on the electrochemical oxidation of NO on a carbon fiber electrode covered with a thin layer of a conducting polymeric metalloporphyrin catalyst, overlaid with another thin film of Nafion, a cation exchange material. The electric current generated during NO oxidation on the surface of the polymeric porphyrin is linearly proportional to the concentration of NO, so this current is used as an analytical signal which can be measured in either the amperometric or the voltammetric mode. Both methods provide a quantitative signal. This unit describes the electrochemical setup for measurement of NO in single cells and tissue. Support protocols describe porphyrin synthesis, sensor preparation, and sensor calibration.
Malinski
T
T
Center of Biomedical Research, Oakland University, Rochester, Michigan, USA.
Huk
I
I
D000818:Animals; D002138:Calibration; D002478:Cells, Cultured; D004563:Electrochemistry / Q000379:methods*; D004867:Equipment Design; D005465:Fluorocarbon Polymers; D007202:Indicators and Reagents; D007475:Ion Exchange Resins; D008832:Microchemistry / Q000379:methods*; D008839:Microelectrodes; D009569:Nitric Oxide / Q000032:analysis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D011166:Porphyrins / Q000737:chemistry*
D005465:Fluorocarbon Polymers; D007202:Indicators and Reagents; D007475:Ion Exchange Resins; D011166:Porphyrins; D009569:Nitric Oxide; C040402:perfluorosulfonic acid; D019001:Nitric Oxide Synthase