Individually Addressable Thin-Film Ultramicroelectrode Array for Spatial Measurements of Single Vesicle Release
Journal article, 2013
Thin-film platinum ultramicroelectrode arrays (MEAs) with subcellular microelectrodes were developed for the spatial measurement of neurotransmitter release across single cells or clusters of single cells. MEAs consisting of 16, 25, and 36 square ultramicroelectrodes with respective widths of 4, 3, and 2 mu m were fabricated on glass substrates by photolithography, thin-film deposition and reactive ion etching. The electrodes in each MEA are tightly defined in a 30 mu m x 30 mu m square, which potentially useful to measure exocytosis across a single cell or clusters of single cells. These MEAs have been characterized with scanning electron microscopy and cyclic voltammetry and show excellent stability and reproducibility. Culturing PC12 cells on top of the MEA.s has been achieved by modifying the array with a poly(dimethylsiloxane) chamber and coating a thin layer of collagen IV top of the electrode surface. The electrochemical response to dopamine has been characterized after coating the surface with the cell-adhering molecules and then with cells attached. Amperometric detection demonstrates that individual exocytotic events can be recorded at these arrays with spatial resolution for dynamic electrochemical measurements near 2 mu m. In contrast to previous single-cell experiments, the effect of dopaminergic drugs on imaging single vesicle exocytotic release from PC 12 cell clusters is presented at cell clusters incubated with the dopamine precursor and Parkinson's therapy agent, L-3,4-dihydroxyphenylalanine, and at cell clusters incubated with the vesicular monoamine transport inhibitor, reserpine. The results of electrochemical imaging demonstrate that the drug effect on PC12 cell clusters is consistent with previous single-cell experiments.
1994
en tk
p554
1995
pheochromocytoma pc12 cells
exocytotic release
v363
quantal catecholamine release
fusion pore
1993
cells
microelectrode arrays
amperometric detection
nature
v66
chronoamperometric
dopamine release
v701
toledo ga
analytical chemistry
response
en gy
p3031
chromaffin
electrodes
brain research
p167
Author
J. Wang
Goteborgs Universitet
R. Trouillon
Goteborgs Universitet
Y. Q. Lin
Goteborgs Universitet
M. I. Svensson
Goteborgs Universitet
Andrew Ewing
Chalmers, Chemical and Biological Engineering, Analytical Chemistry
Analytical Chemistry
0003-2700 (ISSN) 1520-6882 (eISSN)
Vol. 85 11 5600-5608Subject Categories (SSIF 2011)
Analytical Chemistry
DOI
10.1021/ac4009385