Abstract
In this paper, we describe a microchip-based system for amperometrically monitoring the amount of catecholamines released from rat pheochromocytoma (PC 12) cells. Key to this system is a novel, yet simple method for the immobilization of PC 12 cells in poly(dimethylsiloxane) (PDMS)-based microchannels. The procedure involves selectively coating microchannels with collagen followed by introduction of PC 12 cells over the PDMS structure, with the cells being immobilized only on the coated portion of the channels. The cell-coated microchannels can then be reversibly sealed to a glass plate containing electrodes for amperometric detection, resulting in an immobilized cell reactor with integrated microelectrodes. Nafion-coated microelectrodes made by micromolding of carbon inks were used to measure calcium-induced catecholamine release from the cells. Varying concentrations of PC 12 cells immobilized in the microchannels led to a catecholamine release ranging from 20 to 160 μM when the cells were stimulated with a calcium solution. This microchip approach leads to a three-dimensional culture that can be used with this or other cells lines to study the effect of external stimuli on neurotransmitter release.
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