An optimal design method for preventing air bubbles in high-temperature microfluidic devices
An optimal design method for preventing air bubbles in high-temperature microfluidic devices
复制标题
DOI：
10.1007/s00216-009-3160-7
复制
发表时间：
2010-01-01
影响因子：
4.3
通讯作者：
Tamiya, Eiichi
中科院分区：
文献类型：
--
作者：
Nakayama, Tsuyoshi;Hiep, Ha Minh;Tamiya, Eiichi
聚合酶链反应（PCR）的DNA分析已成为医学诊断，环境检查，食品评估和生物学研究的常规部分。此外，微型PCR芯片的开发是旨在将PCR集成到微全分析系统（mu-TAS）中的研究的重要组成部分。然而，微通道中气泡的出现使该过程复杂化。在这项研究中，我们研究了一种新的技术的基础上，层流的流体动力学，利用少量的矿物油在样品注射开始，以防止气泡发生在微通道。我们还进一步优化了压力，加压通道的长度和油的体积，从而使我们的微流控装置更适用于高温PCR。此外，使用优化的PCR芯片进行定量连续PCR，以检测转基因（GM）玉米。从转基因玉米、MON 810和非转基因玉米中提取DNA，浓度从0%（w/v）到100%（w/v）。然后使用基于激光的系统在PCR芯片上分析DNA扩增信号。来自我们的微流控PCR芯片的信号被发现与初始转基因玉米浓度成正比地增加。
DNA analysis with the polymerase chain reaction (PCR) has become a routine part of medical diagnostics, environmental inspections, food evaluations, and biological studies. Furthermore, the development of a microscale PCR chip is an essential component of studies aimed at integrating PCR into a micro total analysis system (mu-TAS). However, the occurrence of air bubbles in microchannels complicates this process. In this study, we investigated a new technique based on the fluid dynamics of laminar flow that utilizes a small amount of mineral oil at the beginning of sample injection to prevent air bubbles from occurring in microchannels. We also further optimized the pressure, the length of the pressurizing channel and the volume of oil, thus making our microfluidic device more useful for high-temperature PCR. Additionally, quantitative continuous-flow PCR was performed using the optimized PCR chip in order to detect genetically modified (GM) maize. DNA was extracted from GM maize, MON 810, and non-GM maize at several concentrations from 0% (w/v) to 100% (w/v). The DNA amplification signals were then analyzed on the PCR chip using a laser-based system. The signal from our microfluidic PCR chip was found to increase in direct proportion to the initial GM maize concentration.