Document Detail


Electroanalysis of metabolic flux from single cells in simple picoliter-volume microsystems.
MedLine Citation:
PMID:  12380823     Owner:  NLM     Status:  PubMed-not-MEDLINE    
Abstract/OtherAbstract:
A picoliter-volume electrochemical analytical chamber has been developed for detecting the metabolic flux resulting from the stress responses of a single plant cell. Electrochemical cells, with volumes as small as 100 pL, were fabricated by controlled electrochemical dissolution of a gold wire sealed in glass (the back-etching of the metal realizing an ultralow-volume titer chamber). In the first instance, the electrode contained within the chamber was characterized by the microinjection of standard aliquots of either ascorbic acid or hydrogen peroxide. In all cases, experimental currents obtained correlated well with theoretical calculations. Subsequently, single plant cells were micromanipulated into the chambers and were exposed to amounts of the detergent SDS (which permeabilized the cell membrane and released the intracellular contents). The flux of metabolite released from a single cell was estimated by using electrochemical-linked assays based upon the enzymes catalase, ascorbate oxidase, and horseradish peroxidase (in each case), in the presence of a mediator. In so doing, we investigated the activity of the cellular protection mechanisms through the determination of peroxides, while the individual cell was "stressed". The technique was found to provide a reliable and reproducible method for making single-cell measurements, using fabrication procedures that are both simple and do not require photolithographic methods.
Authors:
Tomoyuki Yasukawa; Andrew Glidle; Jonathan M Cooper; Tomokazu Matsue
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical chemistry     Volume:  74     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2002 Oct 
Date Detail:
Created Date:  2002-10-16     Completed Date:  2002-11-05     Revised Date:  2003-11-03    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5001-8     Citation Subset:  -    
Affiliation:
Bioelectronics Research Centre, Department of Electronics & Electncal Engineering, University of Glasgow, UK. yasu@elec.gla.ac.uk
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