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PFPE-PEG-PFPE 三嵌段共聚物表面活性剂微滴中的气体串扰及基于液滴微流控技术监测细菌气体代谢。

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

机构信息

Department of Chemistry, Natural Science, Incheon National University, Incheon 22012, Korea.

Research Institute of Basic Sciences, Incheon National University, Incheon 22012, Korea.

出版信息

Biosensors (Basel). 2020 Nov 11;10(11):172. doi: 10.3390/bios10110172.

DOI:10.3390/bios10110172
PMID:33187348
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7697390/
Abstract

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. 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.

摘要

PFPE-PEG-PFPE（全氟聚醚-聚乙二醇-全氟聚醚）表面活性剂已被用于液滴微流控技术中，其具有提供高液滴稳定性和生物相容性的优点。由于该表面活性剂确保了液滴的稳定性，基于液滴的微流控系统已被广泛用于在单分子尺度上封装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由氟化油和 PFPE-PEG-PFPE 表面活性剂形成的液滴之间发生了气体串扰。将含有 LB 肉汤的 K-12 细菌细胞包封在液滴中进行培养，并通过含有酚红的相邻液滴来识别气体串扰。由于细菌在代谢过程中会产生氨气，因此氨气的渗透会导致含有酚红的液滴发生颜色变化。通过反应在包封的液滴内的氯化铵和氢氧化钠也证实了氨气体交换。本文中，我们展示了使用 PFPE-PEG-PFPE 表面活性剂形成液滴时的气体串扰问题，并证实了液滴屏障的密度对气体串扰有影响。我们的结果还表明，基于液滴的微流控技术可以用于通过在培养过程中测定细菌代谢物来监测活细菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66f6/7697390/d37cb3c22bef/biosensors-10-00172-g016.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66f6/7697390/d37cb3c22bef/biosensors-10-00172-g016.jpg

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