Fast and robust sample self-digitization for digital PCR
Fast and robust sample self-digitization for digital PCR
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DOI：
10.1016/j.aca.2020.02.010
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发表时间：
2020-04-22
影响因子：
6.2
通讯作者：
Li, Gang
中科院分区：
文献类型：
--
作者：
Cui, Xu;Wu, Lei;Li, Gang
我们提出了一种简便的样品分割方法，以实现快速和低成本的数字聚合酶链式反应(DPCR)分析。通过使用自数字化(SD)芯片将高百分比的样品体积细分为大量等体积的隔室，该方法可以在几分钟内实现低浪费和高阶样品离散化。SD芯片包含一组用于样品输送的平行微流控通道，每个通道连接两排圆柱孔，以容纳离散的样品。通过利用脱气的PDMS密封板作为可拆卸的真空抽气源，SD芯片自动产生大阵列的小样品体积，而不需要外部抽气和阀门部件。与大多数基于微流体室的样品离散化方法不同，我们的可拆卸SD芯片允许通过空气冲洗离散化样品，然后剥离覆盖的PDMS平板，并用油层密封数字化样品。由于避免了耗时的油洗，这种微流控装置可以更快地实现样品体积的数字化。此外，这种数字化芯片可以分割90%以上的样本体积，这对于可用材料数量较少的应用非常重要。我们还通过将所建议的SD芯片应用于dPCR分析来展示其实用性。(C)2020爱思唯尔B.V.保留所有权利。
We present a facile sample partitioning method to enable rapid and low-cost digital PCR (dPCR) assays. By subdividing a high percentage of the sample volume into a large number of equal volume compartments with a self-digitization (SD) chip, this method can achieve a low-waste and high-order sample discretization in a matter of minutes. The SD chip contains a set of parallel microfluidic channels used for sample delivery, and each channel is connected with two rows of cylindrical wells to hold the discretized sample. By utilizing a degassed PDMS sealing slab as a detachable vacuum pumping source, the SD chip automatically generate large arrays of small sample volumes without requirement of external pumping and valving components. Unlike most microfluidic chamber-based methods for sample discretization, our detachable SD chip allows for discretizing sample with air flushing, then peeling off the cover PDMS slab and sealing the digitized samples with oil layer. Due to obviation of time-consuming oil flushing, such microfluidic device can achieve much faster digitization of sample volumes. Furthermore, this digitization chip can partition more than 90% of a sample volume, which is important for the applications where the amount of material available is small. We also demonstrated the utility of the proposed SD chip by applying it to a dPCR assay. (C) 2020 Elsevier B.V. All rights reserved.