Enzyme Kinetic Measurements Using a Droplet-Based Microfluidic System with a Concentration Gradient
Enzyme Kinetic Measurements Using a Droplet-Based Microfluidic System with a Concentration Gradient
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DOI：
10.1021/ac102472a
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发表时间：
2011-03-01
影响因子：
7.4
通讯作者：
Seong, Gi Hun
中科院分区：
文献类型：
--
作者：
Bui, Minh-Phuong Ngoc;Li, Cheng Ai;Seong, Gi Hun
在本文中，我们提出了一种微流控装置，它能够产生一个浓度梯度，然后在一个简单的T形连接几何形状的通道内形成平行的液滴。基于层流下的流体扩散，可以获得线性浓度梯度分布。使用荧光染料研究了产生线性浓度梯度和平行液滴形成的优化条件。扩散混合下的浓度梯度分布主要由样品入口处的流速决定，而平行液滴的形成受入口和出口处的通道几何形状的影响。微流控装置的实验特征在于使用通过设计过程选择的最佳布局和操作条件。此外，进行试卤灵-β-D-吡喃半乳糖苷的β-半乳糖苷酶催化水解的原位酶动力学测量，以证明我们的简单、时间有效和低样品体积的微流体装置的应用潜力。我们希望，除了酶动力学，药物筛选和临床诊断测试可以快速，准确地使用这种基于液滴的微流体系统进行。
In this Paper We propose a microfluidic device that is capable Of generating a concentration gradient followed by parallel droplet formation within channels with a simple T-junction geometry. Linear concentration gradient profiles can be obtained based on fluid diffusion under laminar flow. Optimized conditions for generating a linear concentration gradient and parallel droplet formation were investigated using fluorescent dye. The concentration gradient profile under diffusive Mixing was dominated by the flow rate at sample inlets, while parallel droplet formation was affected by the channel geometry at both the inlet and outlet The microfluidic device was experimentally characterized using optimal layout and operating conditions,selected through a design process. Furthermore, in situ enzyme kinetic measurements of the beta- galactosidase-catalyzed hydrolysis of resorufin-beta-D-galactopyranoside were,performed to demonstrate the application potential of our Simple, time-effective, and low sample volume microfluidic device. We expect that, in addition to enzyme kinetics, drug screening and clinical diagnostic tests can be rapidly and accurately performed using this droplet-based microfluidic system.