Document Detail


Individually addressable thin film ultra-microelectrode array for spatial measurements of single vesicle release.
MedLine Citation:
PMID:  23627439     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Thin-film platinum ultra-microelectrode arrays (MEAs) with sub-cellular 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 ultra-microelectrodes with respective widths of 4, 3, and 2 µm were fabricated on glass substrates by photolithography, thin-film deposition and reactive ion etching. The electrodes in each MEA are tightly defined in a 30×30 µm square, which is potentially useful to measure exocytosis across 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 MEAs has been achieved by modifying the array with a polydimethysiloxane chamber and coating a thin layer of collagen IV on 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 micrometers. In contrast to previous single cell experiments, the effect of dopaminergic drugs on imaging single vesicle exocytotic release from PC12 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.
Authors:
Jun Wang; Raphael Trouillon; Yuqing Lin; Maria I Svensson; Andrew G Ewing
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-4-29
Journal Detail:
Title:  Analytical chemistry     Volume:  -     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-4-30     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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