Planar chip device for PCR and hybridization with surface acoustic wave pump
Planar chip device for PCR and hybridization with surface acoustic wave pump
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DOI：
10.1039/b412712a
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发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Wixforth, A
中科院分区：
文献类型：
--
作者：
Guttenberg, Z;Müller, H;Wixforth, A
我们开发了一种工作在平面上的微流控器件，而不是闭合的通道网络。利用表面声波(SAW)在压电LiNbO_3衬底上以单个液滴的形式传输流体。压电体的表面具有化学结构，以诱导液滴的高接触角或包围液体可以润湿衬底的区域。将声表面波技术与薄膜电阻加热器相结合，设计了一种集成了DNA聚合酶链式反应和杂交技术的生物分析芯片。为了防止聚合酶链式反应试剂在高温下蒸发，样品被包裹在矿物油液滴中。在这个芯片上，SAW分解干燥的底漆，在杂交过程中将带油的液体在两个加热器之间移动并混合。该芯片能够以低至200nL的体积进行高灵敏、快速和特异的聚合酶链式反应。在温度循环期间，用光学装置在线监测DNA浓度是可行的，提供0.1 ng的灵敏度。在聚合酶链式反应之后，产物被移到第二加热器进行斑点DNA阵列的杂交。利用我们的芯片，我们能够从人类血液中检测到导致莱顿因子V综合征的单核苷酸多态(SNP)。
We have developed a microfluidic device operating at a planar surface instead of a closed channel network. The fluid is transported in single droplets using surface acoustic waves ( SAW) on a piezoelectric LiNbO3 substrate. The surface of the piezo is chemically structured to induce high contact angles of the droplets or enclose areas where the liquid can wet the substrate. Combining the SAW technique with thin film resistance heaters, a biological analysis chip with integrated DNA amplification by PCR and hybridization was designed. To prevent evaporation of the PCR reagents at high temperatures the sample is enclosed in droplets of mineral oil. On this chip the SAW resolves dried primers, shifts the oil capped liquid between the two heaters and mixes during hybridization. The chip is able to perform a highly sensitive, fast and specific PCR with a volume as low as 200 nl. During the temperature cycles an online monitoring of the DNA concentration is feasible with an optical unit, providing a sensitivity of 0.1 ng. After PCR the product is moved to the second heater for the hybridization on a spotted DNA array. With our chip we were able to detect a single nucleotide polymorphism ( SNP) responsible for the Leiden Factor V syndrome from human blood.