A Microchip-Based System for Immobilizing PC 12 Cells and Amperometrically Detecting Catecholamines Released After Stimulation with Calcium

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 mM 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.

A compact multifunctional microfluidic platform for exploring cellular dynamics in real-time using electrochemical detection

“…[39][40][41][42][43] Our results indicate that when chemical stimulation is to be used it is signicant to optimise the ow conditions in order to avoid mechanical stimulation, however, this aspect is not commonly addressed in publications.…”

Biomimetic Sensors for Rapid Testing of Water Resources

“…By harnessing the intrinsic potential of certain cells to form aligned tissues in vivo, we can generate functional tissues in vitro by confining the cells in the appropriate 3D microarchitecture [30]. Cells possessing the prerequisite ability to self-organize in vivo include fibroblasts, myoblasts, endothelial cells, and cardiac stem cells.…”

On‐Chip Determination of Dopamine Exocytosis Using Mercaptopropionic Acid Modified Microelectrodes

“…Measurement of adrenaline and noradrenalin exocytosis from Chromaffin cells was demonstrated by Chen et al in a gold plated picoliter well [11], by Sun et al in a microfluidic system containing a transparent ITO electrode [12] while Dias et al used a Pt array on glass [13]. DA exocytosis from PC12 cells has been measured in a microfluidic system containing a Pt-electrode [14] whereas Huang et al used a microfluidic system to position a cell prior to measurement using a carbon fiber microelectrode [5].…”