Austin C M, Caro D M, Sankar S, Penniman W F, Perdomo J E, Hu L, Patel S, Gu X, Watve S, Hammer B K, Forest C R
Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
Massachusetts Institute of Technology, Cambridge Massachusetts 02139, USA.
Biomicrofluidics. 2017 Jul 31;11(4):044110. doi: 10.1063/1.4995597. eCollection 2017 Jul.
Genetically engineered bacteria can be used for a wide range of applications, from monitoring environmental toxins to studying complex communication networks in the human digestive system. Although great strides have been made in studying single strains of bacteria in well-controlled microfluidic environments, there remains a need for tools to reliably control and measure communication between multiple discrete bacterial populations. Stable long-term experiments (e.g., days) with controlled population sizes and regulated input (e.g., concentration) and output measurements can reveal fundamental limits of cell-to-cell communication. In this work, we developed a microfluidic platform that utilizes a porous monolith to reliably and stably partition adjacent strains of bacteria while allowing molecular communication between them for several days. We measured small molecule production by the bacterial populations in response to stimuli using analytical chemistry methods and measured fluorescent output. The results are compared with communication and diffusion delay models. This porous monolith microfluidic system enables bacterial cell-to-cell communication assays with dynamic control of inputs, relatively long-term experimentation with no cross contamination, and stable bacterial population size. This system can serve as a valuable tool in understanding bacterial communication and improving biosensor design capabilities.
基因工程细菌可用于广泛的应用，从监测环境毒素到研究人类消化系统中的复杂通信网络。尽管在精心控制的微流体环境中研究单一菌株的细菌方面已经取得了很大进展，但仍然需要可靠地控制和测量多个离散细菌群体之间通信的工具。具有受控种群大小、调节输入（例如浓度）和输出测量的稳定长期实验（例如数天）可以揭示细胞间通信的基本限制。在这项工作中，我们开发了一种微流体平台，该平台利用多孔整体柱可靠且稳定地分隔相邻的细菌菌株，同时允许它们之间进行数天的分子通信。我们使用分析化学方法测量了细菌群体对刺激的小分子产生，并测量了荧光输出。将结果与通信和扩散延迟模型进行了比较。这种多孔整体柱微流体系统能够进行细菌细胞间通信分析，对输入进行动态控制，进行无交叉污染的相对长期实验，并保持稳定的细菌种群大小。该系统可作为理解细菌通信和提高生物传感器设计能力的有价值工具。
