Water-in-oil microdroplets offer microreactors for compartmentalized biochemical reactions with high throughput. Recently, the combination with a sol-gel switch ability, using agarose-in-oil microdroplets, has increased the range of possible applications, allowing for example the capture of amplicons in the gel phase for the preservation of monoclonality during a PCR reaction. Here, we report a new method for generating such agarose-in-oil microdroplets on a microfluidic device, with minimized inlet dead volume, on-chip cooling, and in situ monitoring of biochemical reactions within the gelified microbeads. We used a flow-focusing microchannel network and successfully generated agarose microdroplets at room temperature using the “push-pull” method. This method consists in pushing the oil continuous phase only, while suction is applied to the device outlet. The agarose phase present at the inlet is thus aspirated in the device, and segmented in microdroplets. The cooling system consists of two copper wires embedded in the microfluidic device. The transition from agarose microdroplets to microbeads provides additional stability and facilitated manipulation. We demonstrate the potential of this method by performing on-chip a temperature-triggered DNA isothermal amplification in agarose microbeads. Our device thus provides a new way to generate microbeads with high throughput and no dead volume for biochemical applications.
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December 2012
Research Article|
October 09 2012
A microfluidic device for on-chip agarose microbead generation with ultralow reagent consumption
Linda Desbois;
Linda Desbois
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Adrien Padirac;
Adrien Padirac
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Shohei Kaneda;
Shohei Kaneda
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Anthony J. Genot;
Anthony J. Genot
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Yannick Rondelez;
Yannick Rondelez
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Didier Hober;
Didier Hober
2
Université Lille 2, Faculté de Médecine et CHRU Lille
, Laboratoire de Virologie/EA 3610, Loos-lez-lille 59120, France
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Dominique Collard;
Dominique Collard
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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Teruo Fujii
Teruo Fujii
a)
1LIMMS/CNRS-IIS, Institute of Industrial Science,
The University of Tokyo
, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
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a)
Author to whom correspondence should be addressed. Electronic mail: tfujii@iis.u-tokyo.ac.jp.
Biomicrofluidics 6, 044101 (2012)
Article history
Received:
July 23 2012
Accepted:
September 27 2012
Citation
Linda Desbois, Adrien Padirac, Shohei Kaneda, Anthony J. Genot, Yannick Rondelez, Didier Hober, Dominique Collard, Teruo Fujii; A microfluidic device for on-chip agarose microbead generation with ultralow reagent consumption. Biomicrofluidics 1 December 2012; 6 (4): 044101. https://doi.org/10.1063/1.4758460
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