Microfluidic long-term differential oxygenation for bacterial growth characteristics analyses
Microfluidic long-term differential oxygenation for bacterial growth characteristics analyses
复制标题
DOI：
10.1039/c4ra01577k
复制
发表时间：
2014-04
期刊：
影响因子：
3.9
通讯作者：
Xin Cui;H. Yip;Qian Zhu;Cheng-Wei Yang;Raymond H. W. Lam
中科院分区：
文献类型：
--
作者：
Xin Cui;H. Yip;Qian Zhu;Cheng-Wei Yang;Raymond H. W. Lam
溶解氧是决定细菌生长特性（如细胞活力、迁移、聚集和代谢过程）的关键微环境因素。在这里，我们报告了一种微流体长期充氧器，它可以在多种定义的充氧条件（0-42 ppm）下支持高通量平行细胞培养。该装置中开发并制造了一系列氧气-氮气微混合器，为氧化过程产生稳定的氧气浓度。位于气体层和细胞培养室之间的水套层用于阻止蒸发并维持室中培养基的条件。此外，我们还进行模拟和实验来研究气体混合器和水套的功能。我们还进行培养实验，研究三种选定的牙科细菌（变形链球菌、粘性放线菌和具核梭杆菌）在一定氧合条件下的长期生长（长达一周）和聚集。这些特殊的结果可以为了解它们在牙齿生物膜形成中的作用提供重要的见解。总的来说，这项工作表明，长期微流体充氧方法可以有效调节细胞微环境中规定的溶解氧水平。重要的是，这种氧合方法可以进一步应用于细胞行为、代谢和遗传反应及其生物膜形成过程的一般长期分析。
Dissolved oxygen is a critical micro-environmental factor to determine the growth characteristics of bacteria, such as cell viability, migration, aggregation and metabolic processes. Here, we report a microfluidic long-term oxygenator which can support high-throughput parallel cell cultures under multiple defined oxygenation conditions (0–42 ppm). An array of oxygen–nitrogen gas micro-mixers is developed and fabricated in the device to generate stable oxygen concentrations for the oxygenation process. A water-jacket layer located between the gas layer and the cell culture chamber is applied to block evaporation and maintain the conditions of the culture media in the chamber. Furthermore, we perform simulations and experiments to investigate the functions of the gas mixers and the water jackets. We also conduct culture experiments to study the long-term growth (up to one week) and aggregation of three selected dental bacteria (Streptococcus mutans, Actinomyces viscosus and Fusobacterium nucleatum) under ranges of oxygenation conditions. These particular results can provide important insights into their roles in dental biofilm formation. Overall, this work demonstrates that the long-term microfluidic oxygenation approach can effectively regulate defined dissolved-oxygen levels in cell microenvironments. Importantly, this oxygenation approach can be further applied to general long-term analyses of cells for their behavioral, metabolic and genetic responses, and their biofilm formation processes.