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Gas Crosstalk between PFPE–PEG–PFPE Triblock Copolymer Surfactant-Based Microdroplets and Monitoring Bacterial Gas Metabolism with Droplet-Based Microfluidics

机译:PFPE-PEG-PFPE三嵌段共聚物表面活性剂的微量轧体和监测基于液滴的微流体的细菌气体代谢之间的气体串扰

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摘要

The PFPE–PEG–PFPE (Perfluoropolyether-polyethylene glycol-perfluoropolyether) surfactant has been used in droplet-based microfluidics and is known to provide high droplet stability and biocompatibility. Since this surfactant ensures the stability of droplets, droplet-based microfluidic systems have been widely used to encapsulate and analyze various biological components at the single-molecule scale, including viruses, bacteria, nucleic acids and proteins. In this study, we experimentally confirmed that gas crosstalk occurred between droplets formed by fluorinated oil and the PFPE–PEG–PFPE surfactant. E. coli K-12 bacterial cells were encapsulated with Luria–Bertani broth within droplets for the cultivation, and gas crosstalk was identified with neighboring droplets that contain phenol red. Since bacteria produce ammonia gas during its metabolism, penetration of ammonia gas initiates a color change of phenol red-containing droplets. Ammonia gas exchange was also confirmed by reacting ammonium chloride and sodium hydroxide within droplets that encapsulated. Herein, we demonstrate the gas crosstalk issue between droplets when it is formed using the PFPE–PEG–PFPE surfactant and also confirm that the density of droplet barrier has effects on gas crosstalk. Our results also suggest that droplet-based microfluidics can be used for the monitoring of living bacteria by the determination of bacterial metabolites during cultivation.
机译:PFPE-PEG-PFPE(全氟聚醚 - 聚乙二醇 - 全氟聚醚)表面活性剂已用于液滴基微流体,并已知提供高液滴稳定性和生物相容性。由于该表面活性剂确保了液滴的稳定性,因此基于液滴的微流体系统已被广泛用于在单分子规模处包封和分析各种生物成分,包括病毒,细菌,核酸和蛋白质。在这项研究中,我们通过实验证实了通过氟化油和PFPE-PEG-PFPE表面活性剂形成的液滴之间发生气体串扰。将大肠杆菌K-12细菌细胞用Luria-Bertani肉汤包封在栽培中的液滴内,并用含有苯酚红色的相邻液滴鉴定气体串扰。由于细菌在代谢过程中产生氨气,因此氨气的渗透引起含酚红色液滴的颜色变化。还通过将氯化铵和氢氧化钠在包封的液滴内反应来证实氨气交换。在此,我们在使用PFPE-PEG-PFPE表面活性剂形成时展示液滴之间的气体串扰问题,并且还证实液滴屏障的密度对气体串扰产生影响。我们的研究结果还表明,基于液滴的微流体可用于通过测定培养过程中的细菌代谢物来监测活细菌。

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