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Hands-Off Preparation of Monodisperse Emulsion Droplets Using a Poly(dimethylsiloxane) Microfluidic Chip for Droplet Digital PCR

机译:使用聚二甲基硅氧烷微流控芯片进行液滴数字PCR的单分散乳液液滴的手动制备

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摘要

A fully autonomous method of creating highly monodispersed emulsion droplets with a low sample dead volume was realized using a degassed poly(dimethylsiloxane) (PDMS) microfluidic chip possessing a simple T-junction channel geometry with two inlet reservoirs for oil and water to be loaded and one outlet reservoir for the collection of generated droplets. Autonomous transport of oil and water phases in the channel was executed by permeation of air confined inside the outlet reservoir into the degassed PDMS. The only operation required for droplet creation was simple pipetting of oil and aqueous solutions into the inlet reservoirs. Long-lasting fluid transport in the current system enabled us to create ca. 51,000 monodispersed droplets (with a coefficient of variation of <3% for the droplet diameter) in 80 min with a maximum droplet generation rate of ca. 12 Hz using a PDMS chip that had been degassed overnight. With multiple time-course measurements, the reproducibility in the current method of droplet preparation was confirmed, with tunable droplet sizes achieved simply by changing the cross-sectional dimensions of the microchannel. Furthermore, it was verified that the resultant droplets could serve as microreactors for digital polymerase chain reactions. This hands-free technique for preparing monodispersed droplets in a very facile and inexpensive fashion is intended for, but not limited to, bioanalytical applications and is also applicable to material syntheses.
机译:使用脱气的聚二甲基硅氧烷(PDMS)微流控芯片实现了一种完全自动化的方法,该方法可以创建具有低样品死体积的高度单分散的乳液液滴,该芯片具有简单的T型结通道几何结构,带有两个用于装载油和水的入口储油罐,并且一个出口容器,用于收集产生的液滴。通道中油相和水相的自主运输是通过将出口储油罐内的空气渗透到脱气的PDMS中来实现的。产生液滴所需的唯一操作是将油和水溶液简单地移液到入口容器中。当前系统中的流体长期传输使我们能够创建大约80分钟内有51,000个单分散液滴(液滴直径的变化系数<3%),最大液滴产生速率约为。使用已脱气过夜的PDMS芯片,频率为12 Hz。通过多次时程测量,可以确定当前液滴制备方法的可重复性,并且只需更改微通道的横截面尺寸即可实现可调节的液滴尺寸。此外,已证实所得的液滴可用作数字聚合酶链反应的微反应器。这种以非常容易和廉价的方式制备单分散液滴的免提技术旨在但不限于生物分析应用,并且还适用于材料合成。

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