Aliquoting on the centrifugal microfluidic platform based on centrifugo-pneumatic valves
Aliquoting on the centrifugal microfluidic platform based on centrifugo-pneumatic valves
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DOI：
10.1007/s10404-010-0759-0
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发表时间：
2011-06-01
影响因子：
2.8
通讯作者：
von Stetten, Felix
中科院分区：
文献类型：
--
作者：
Mark, Daniel;Weber, Patrick;von Stetten, Felix
提出了一种在离心微流控平台上进行液体分装的新方法。等分是从单个样品进行多个平行测定（“几何多重”）的基本单元操作，例如通过实时聚合酶链反应（PCR）或均相免疫测定板进行基因分型。我们的方法是一个两阶段的过程与初始计量阶段和随后的运输阶段开始切换离心气动阀。该方法能够将液体等分到完全分离的反应腔中。它包括独立于接收腔中预存试剂体积的精确计量。它还排除了接收腔之间的任何交叉污染。我们的特点是三种不同的技术制造的原型的性能：微铣削，热成型箔，注塑成型。将类似于90 μ l的初始体积分成8个等份，每个等份10 μ l体积，加上热成型箔盘上的废物储存器，导致计量体积的变异系数（CV）为3.6%。将类似于105 μ l的类似体积分成16个等份，每个等份6 μ l体积，置于微研磨和注塑盘上，相应的CV分别为2.8和2.2%。因此，证明了新颖的等分结构与上述原型和生产技术的兼容性。此外，实验和理论上解决了等分结构相对于这些生产技术中的每一个所固有的生产公差的可实现的体积精度的重要问题。这种新方法适合低成本大规模生产，因为它不需要任何复制后的表面修饰，如疏水补丁。
We present a new method for aliquoting liquids on the centrifugal microfluidic platform. Aliquoting is an essential unit operation to perform multiple parallel assays ("geometric multiplexing") from one individual sample, such as genotyping by real-time polymerase chain reactions (PCR), or homogeneous immunoassay panels. Our method is a two-stage process with an initial metering phase and a subsequent transport phase initiated by switching a centrifugo-pneumatic valve. The method enables aliquoting liquids into completely separated reaction cavities. It includes precise metering that is independent on the volume of pre-stored reagents in the receiving cavities. It further excludes any cross-contamination between the receiving cavities. We characterized the performance for prototypes fabricated by three different technologies: micro-milling, thermoforming of foils, and injection molding. An initial volume of similar to 90 mu l was split into 8 aliquots of 10 mu l volume each plus a waste reservoir on a thermoformed foil disk resulting in a coefficient of variation (CV) of the metered volumes of 3.6%. A similar volume of similar to 105 mu l was split into 16 aliquots of 6 mu l volume each on micro-milled and injection-molded disks and the corresponding CVs were 2.8 and 2.2%, respectively. Thus, the compatibility of the novel aliquoting structure to the aforementioned prototyping and production technologies is demonstrated. Additionally, the important question of achievable volume precision of the aliquoting structure with respect to the production tolerances inherent to each of these production technologies is addressed experimentally and theoretically. The new method is amenable to low cost mass production, since it does not require any post-replication surface modifications like hydrophobic patches.