Department of Biochemistry, University of Cambridge, Cambridge, CB2 1GA, United Kingdom.
J Am Chem Soc. 2009 Oct 28;131(42):15251-6. doi: 10.1021/ja904823z.
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.
已经在 PDMS 中制造出一种能够以恒定体积存储包含单个细菌的皮升级液滴的微流控设备。一旦被捕获在设备中保持静态的液滴中，细菌就会从双顺反子构建体中表达红色荧光蛋白（mRFP1）和碱性磷酸酶（AP）两种酶。通过测量细胞内 mRFP1 的荧光强度和细胞外酶活性产生的荧光产物的强度，可以同时连续监测基因表达和酶活性。通过从许多单个细胞收集数据，可以定量细胞内活性的分布，并测量两种 AP 突变体之间的活性差异。
