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Issue 1, 2005
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Perfusion and chemical monitoring of living cells on a microfluidic chip

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Abstract

A microfluidic device that incorporates continuous perfusion and an on-line electrophoresis immunoassay was developed, characterized, and applied to monitoring insulin secretion from single islets of Langerhans. In the device, a cell chamber was perfused with cell culture media or a balanced salt solution at 0.6 to 1.5 µL min−1. The flow was driven by gas pressure applied off-chip. Perfusate was continuously sampled at 2 nL min−1 by electroosmosis through a separate channel on the chip. The perfusate was mixed on-line with fluorescein isothiocyanate-labeled insulin (FITC-insulin) and monoclonal anti-insulin antibody and allowed to react for 60 s as the mixture traveled down a 4 cm long reaction channel. The cell chamber and reaction channel were maintained at 37 °C. The reaction mixture was injected onto a 1.5 cm separation channel as rapidly as every 6 s, and the free FITC-insulin and the FITC-insulin-antibody complex were separated under an electric field of 500 to 600 V cm−1. The immunoassay had a detection limit of 0.8 nM and a relative standard deviation of 6% during 2 h of continuous operation with standard solutions. Individual islets were monitored for up to 1 h while perfusing with different concentrations of glucose. The immunoassay allowed quantitative monitoring of classical biphasic and oscillatory insulin secretion with 6 s sampling frequency following step changes in glucose from 3 to 11 mM. The 2.5 cm × 7.6 cm microfluidic system allowed for monitoring islets in a highly automated fashion. The technique should be amenable to studies involving other tissues or cells that release chemicals.

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Publication details

The article was received on 02 Apr 2004, accepted on 20 May 2004 and first published on 22 Jul 2004


Article type: Paper
DOI: 10.1039/B404974H
Citation: Lab Chip, 2005,5, 56-63
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    Perfusion and chemical monitoring of living cells on a microfluidic chip

    J. G. Shackman, G. M. Dahlgren, J. L. Peters and R. T. Kennedy, Lab Chip, 2005, 5, 56
    DOI: 10.1039/B404974H

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