Related Experiment Video
Updated: Mar 7, 2026
09:26
High-throughput Protein Expression Generator Using a Microfluidic Platform
Published on: August 23, 2012
12.3K
High-throughput screening of microchip-synthesized genes in programmable double-emulsion droplets
1Department of Biomedical Engineering, Duke University, Durham, 27705, USA. kam.leong@columbia.edu and Department of Biomedical Engineering, Columbia University, New York, 10027, USA.
Nanoscale
|February 28, 2017
Summary
This study introduces a novel microfluidic platform using double emulsion droplets for high-throughput screening of synthetic genes. The technology enhances fluorescence signals for efficient sorting of functional gene variants in Escherichia coli.
Area of Science:
- Synthetic biology
- Biotechnology
- Microfluidics
- High-throughput screening
Background:
- Synthetic biology and biotechnology require advanced high-throughput screening methods for optimizing protein expression.
- Current single-emulsion droplet systems face limitations in aqueous phase analysis and inter-phase communication.
- Microfluidic devices offer potential for droplet-based high-throughput screening.
Purpose of the Study:
- To develop a high-throughput, miniaturized screening platform for microchip-synthesized genes.
- To overcome limitations of single-emulsion droplets using water-in-oil-in-water (W/O/W) double emulsion (DE) droplets.
- To enable efficient screening and enrichment of functional synthetic gene variants.
Main Methods:
- Utilized microfluidics to generate W/O/W double emulsion (DE) droplets.
- Synthesized synthetic gene variants of fluorescent proteins using a microarray inkjet synthesizer.
- Encapsulated single Escherichia coli (E. coli) cells with gene variants into DE droplets for screening.
- Induced gene expression using isopropyl β-d-1-thiogalactopyranoside (IPTG) permeation through the oil layer.
- Employed fluorescence-activated cell sorting (FACS) for droplet analysis.
Main Results:
- Achieved a 100-fold enhancement in fluorescence signals within 24 hours of bacterial proliferation in DE droplets.
- Demonstrated enrichment of functional gene variants, including the red fluorescent protein (rfp) gene, using an error correction method.
- Generated detectable fluorescence signals from approximately 100 bacteria per droplet, enabling FACS.
- Successfully screened microchip-synthesized genes and obviated the need for conventional bulk cell culture.
Conclusions:
- The developed DE droplet platform provides a powerful tool for high-throughput screening of synthetic genes.
- This technology enhances signal detection and enables efficient sorting of functional gene variants.
- The platform overcomes previous limitations, offering a more efficient and less labor-intensive approach for synthetic biology applications.
