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A Thermosetting Oil for Droplet-Based Real-Time Monitoring of Digital PCR and Cell Culture

机译：一种热固性油，用于基于液滴的实时数字PCR和细胞培养监测

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摘要

Real-time observation of small volumes helps to understand phenomena ranging from reaction kinetics to cellular heterogeneity. However, high-throughput real-time monitoring systems based on droplets are rarely reported due to droplet motion and coalescence. Herein, a thermosetting oil is developed as a continuous phase to solve these problems through transforming into elastic solid after droplet generation. The thermosetting oil consists of silicon oil, nonionic surfactant, vinyl silicone oil, hydrosilicone oil, and Pt catalyst. Using Pt as the catalyst, the polymerization of this mixture can be accomplished in 72 min and 37 s at 37 and 95 degrees C, respectively, based on the hydrosilation of vinyl silicone oil and hydrosilicone oil without other inducers (e.g., UV, Ca2+). Droplet-based real-time monitoring of digital polymerase chain reaction (PCR) and cell culture is successfully achieved using this thermosetting oil, demonstrating that droplets motion and coalescence can be eliminated by the crosslinking material without the influence on bioreactions. In addition, the PCR products can be totally recovered from microcapsules created by this oil for further analysis. Thus, the newly developed oil allows real-timely monitoring individual droplets in a high-throughput manner, as well as has potential for other bioreactions and cell-based assays.
机译：实时观察小体积有助于理解从反应动力学到细胞异质性的现象。然而，由于液滴运动和聚结，很少有基于液滴的高通量实时监测系统的报道。在本文中，开发了热固性油作为连续相以通过在液滴产生后转化成弹性固体来解决这些问题。热固性油由硅油，非离子表面活性剂，乙烯基硅油，氢硅油和Pt催化剂组成。使用Pt作为催化剂，基于乙烯基硅油和氢硅油的硅氢化反应而无需其他诱导剂（例如UV，Ca2 +），可以分别在37和95摄氏度下分别在72分钟和37秒内完成该混合物的聚合。 。使用这种热固性油成功地实现了基于液滴的数字聚合酶链反应（PCR）和细胞培养的实时监控，这表明交联材料可以消除液滴的运动和聚结，而不会影响生物反应。此外，PCR产物可从该油产生的微囊中完全回收，以进行进一步分析。因此，新开发的油可以以高通量的方式实时监测单个液滴，并且具有进行其他生物反应和基于细胞的测定的潜力。

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  • 来源
    《Advanced Functional Materials》 |2018年第39期|1803559.1-1803559.11|共11页
  • 作者

    Wu Wenshuai; Zhou Shufang; Hu Jiumei; Wang Guoping; Ding Xiong; Gou Tong; Sun Jingjing; Zhang Tao; Mu Ying;

  • 作者单位

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

    Zhejiang Univ, State Key Lab Ind Control Technol, Inst Cyber Syst & Control, Res Ctr Analyt Instrumentat, Hangzhou 310027, Zhejiang, Peoples R China;

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  • 正文语种 eng
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  • 关键词

    droplet digital PCR; immobile droplets array; real-time monitoring; single cell culture; thermosetting materials;

    机译：液滴数字PCR;固定液滴阵列;实时监测;单细胞培养;热固性材料;

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