A double-emulsion microfluidic platform for in vitro green fluorescent protein expression

Wu, N, Oakeshott, J, Easton, C, Peat, T, Surjadi, R and Zhu, Y 2011, 'A double-emulsion microfluidic platform for in vitro green fluorescent protein expression', Journal of Micromechanics and Microengineering, vol. 21, no. 5, pp. 1-7.

Document type: Journal Article
Collection: Journal Articles

Title A double-emulsion microfluidic platform for in vitro green fluorescent protein expression
Author(s) Wu, N
Oakeshott, J
Easton, C
Peat, T
Surjadi, R
Zhu, Y
Year 2011
Journal name Journal of Micromechanics and Microengineering
Volume number 21
Issue number 5
Start page 1
End page 7
Total pages 7
Publisher Institute of Physics Publishing
Abstract Microfluidic droplet technology has gained popularity due to the advantages over conventional emulsion techniques and capabilities for a wide range of applications. In this paper, the development of a simple microfluidic-based double-emulsion system is reported. Such a system could be potentially used for in vitro protein synthesis. The system involves a two-step process to make water-in-oil-in-water (W/O/W) emulsions. A PMMA microchip is used for the formation of water-in-oil (W/O) single-emulsion droplets. Then, the single-emulsion droplets are transported to a PDMS/glass microchip to make the W/O/W double-emulsion droplets. The system was first characterized by detecting fluorescein sodium salt as a model dye in the internal aqueous droplets using laser-induced fluorescence. The effect of the flow rates of the internal aqueous phase and outer continuous aqueous phase on the formation of the double-emulsion droplets is investigated to provide information for system optimization. On-chip storage of double-emulsion droplets is also investigated to allow for protein synthesis from a PCR-generated DNA template using either commercial in vitro transcription and translation kits or crude Escherichia coli S30 extracts. In vitro expression of the green fluorescent protein is successfully demonstrated in this system.
Subject Manufacturing Engineering not elsewhere classified
Keyword(s) Monodisperse Double Emulsions
Droplet Microfluidics
Single Cells
DOI - identifier 10.1088/0960-1317/21/5/054032
Copyright notice © 2011 IOP Publishing Ltd.
ISSN 0960-1317
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