A microphysiometer for simultaneous measurement of changes in extracellular glucose, lactate, oxygen, and acidification rate

Abstract

A microphysiometer capable of measuring changes in extracellular glucose, lactate, oxygen, and acidification rate has been developed by incorporating modified electrodes into a standard Cytosensor Microphysiometer plunger. Glucose and lactate are measured indirectly at platinum electrodes by amperometric oxidation of hydrogen peroxide, which is produced from catalysis of glucose and lactate at films containing their respective entrapped oxidase. Oxygen is measured amperometrically at a platinum electrode coated with a Nafion film, while the acidification rate is measured potentiometrically by a Cytosensor Microphysiometer. Analytical information is obtained during the Cytosensor stop-flow cycles, where the electrodes measure changes in the extracellular medium corresponding to the consumption or production of the analyte by the cells. Modification of the Cytosensor plunger for multianalyte determination is described, and the operation of the technique is illustrated by the simultaneous measurement of all four analytes during the addition of fluoride and DNP to Chinese hamster ovary cells and fluoride and antimycin A to mouse fibroblast cells. Cell metabolic recovery and dynamics after exposure to agents can also be observed in specific cases.