Format

Send to

Choose Destination
See comment in PubMed Commons below
Biosens Bioelectron. 2010 Jan 15;25(5):1019-24. doi: 10.1016/j.bios.2009.09.018. Epub 2009 Sep 19.

Enzyme electrodes to monitor glucose consumption of single cardiac myocytes in sub-nanoliter volumes.

Author information

1
Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37235-1631, USA.

Abstract

Monitoring the metabolic activity of cells in automated culture systems is one of the key features of micro-total-analysis-systems. We have developed a microfluidic device that allows us to trap single cardiac myocytes (SCMs) in sub-nanoliter volumes and incorporate amperometric glucose-sensing electrodes with working areas of 0.002 mm(2) to measure the glucose consumption of SCM. The miniaturized planar glucose electrodes were fabricated by spin coating platinum electrodes on glass substrates with a glutaraldehyde/enzyme solution and a protective Nafion membrane. The glucose electrodes demonstrate a high enzymatic activity characterized by an apparent Michaelis-Menten constant of 7.52+/-0.18 mM and a sensitivity of approximately 33.8 and approximately 13.2 mA/Mcm(2) at glucose concentration from 0-6 to 6-20 mM in Tyrode's solution, respectively. The response time of the glucose electrodes was between 5 and 15s, and the sensitivity of the electrodes did not degrade over a period of 8 weeks. A replica molded polydimethylsiloxane microfluidic device with a sub-nanoliter sensing volume was sealed to the glass substrate and aligned with the glucose microelectrodes. SCM can be trapped in the sensing volume above the glucose electrodes to measure the glucose consumption over time. The average glucose consumption of SCM was 0.211+/-0.097 mM/min (n=7) in Tyrode's solution with 5 mM of glucose.

PMID:
19833499
PMCID:
PMC4119477
DOI:
10.1016/j.bios.2009.09.018
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Support Center