Continuous flow microfluidic device for cell separation, cell lysis and DNA purification
Continuous flow microfluidic device for cell separation, cell lysis and DNA purification
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DOI：
10.1016/j.aca.2006.11.057
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发表时间：
2007-02-19
影响因子：
6.2
通讯作者：
Chen, Jian
中科院分区：
文献类型：
--
作者：
Chen, Xing;Cui, Dafu;Chen, Jian
研制了一种由微过滤器、微混合器、微柱阵列、微堰、微通道、微腔室和多孔基质组成的新型集成微流控装置，用于全血样品的前处理。细胞分离、细胞裂解和DNA提纯是在这个微型设备中进行的连续流动过程。交叉流过滤用于分离血细胞，可以成功地避免堵塞或堵塞。血细胞在胍缓冲液中裂解后，白细胞中的基因组DNA被释放并吸附在由氢氟酸/乙醇电解液中的硅阳极氧化制备的多孔基质上。对溶液的流动过程进行了模拟和优化。对多孔基质的阳极氧化工艺进行了研究。采用细胞分离、细胞裂解和DNA吸附的连续流程，在集成微流控装置上从1亩L大鼠全血中平均纯化了35.7ngDNA。与商业离心机方法相比，该微型设备可以在50分钟内提取等量的可扩增的DNA。这种集成的微型化设备的最大潜力通过对全血样本的前处理得到说明，最终将样本制备、聚合酶链式反应和分离集成在一个设备上，可能使在护理点基因分析、环境测试和生物战剂检测领域实现完整的检测。(C)2006爱思唯尔B.V.保留所有权利。
A novel integrated microfluidic device that consisted of microfilter, micromixer, micropillar array, microweir, microchannel, microchamber, and porous matrix was developed to perform sample pre-treatment of whole blood. Cell separation, cell lysis and DNA purification were performed in this miniaturized device during a continuous flow process. Crossflow filtration was proposed to separate blood cells, which could successfully avoid clogging or jamming. After blood cells were lyzed in guanidine buffer, genomic DNA in white blood cells was released and adsorbed on porous matrix fabricated by anodizing silicon in HF/ethanol electrolyte. The flow process of solutions was simulated and optimized. The anodization process of porous matrix was also studied. Using the continuous flow procedure of cell separation, cell lysis and DNA adsorption, average 35.7 ng genomic DNA was purified on the integrated microfluidic device from 1 mu L rat whole blood. Comparison with a commercial centrifuge method, the miniaturized device can extract comparable amounts of PCR-amplifiable DNA in 50 min. The greatest potential of this integrated miniaturized device was illustrated by pre-treating whole blood sample, where eventual integration of sample preparation, PCR, and separation on a single device could potentially enable complete detection in the fields of point-of-care genetic analysis, environmental testing, and biological warfare agent detection. (c) 2006 Elsevier B.V. All rights reserved.