Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 131, Issue 42, Pages 15251-15256Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ja904823z
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Funding
- RCUK Basic Technology
- EU NEST MiFem
- EPSRC
- Glaxo Smith Kline
- EPSRC [EP/D048664/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/D048664/1] Funding Source: researchfish
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A microfluidic device capable of storing picoliter droplets containing single bacteria at constant volumes has been fabricated in PDMS. Once captured in droplets that remain static in the device, bacteria express both a red fluorescent protein (mRFP1) and the enzyme, alkaline phosphatase (AP), from a biscistronic construct. By measuring the fluorescence intensity of both the mRFP1 inside the cells and a fluorescent product formed as a result of the enzymatic activity outside the cells, gene expression and enzymatic activity can be simultaneously and continuously monitored. By collecting data from many individual cells, the distribution of activities in a cell is quantified and the difference in activity between two AP mutants is measured.
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