Zhang Yingdan, Zhao Jingru, Cheng Hang, Wang Jing, Yang Liang, Liang Haihua
School of Medicine, Southern University of Science and Technology, Shenzhen, China.
College of Life Sciences, Northwest University, Xi'an, ShaanXi, China.
Bio Protoc. 2021 Aug 20;11(16):e4126. doi: 10.21769/BioProtoc.4126.
Characterization of biofilm formation and metabolic activities is critical to investigating biofilm interactions with environmental factors and illustrating biofilm regulatory mechanisms. An appropriate model that mimics biofilm habitats therefore demands accurate quantitation and investigation of biofilm-associated activities. Current methodologies commonly involve static biofilm setups (such as biofilm assays in microplates, bead biofilms, or biofilms on glass-slides) and fluidic flow biofilm systems (such as drip-flow biofilm reactors, 3-channel biofilm reactors, or tubing biofilm reactors). Continuous flow systems take into consideration the contribution of hydrodynamic shear forces, nutrient supply, and physical transport of dispersed cells, which define the habitat for biofilm development in most natural and engineered systems. This protocol describes the assembly of 3 flow-system setups to cultivate PAO1 and MR-1 model biofilms, including the respective quantitation and observation approaches. The standardized flow systems promise productive and reproducible biofilm experimental results, which can be further modified according to specific research projects.
生物膜形成和代谢活动的表征对于研究生物膜与环境因素的相互作用以及阐明生物膜调控机制至关重要。因此，一个模拟生物膜栖息地的合适模型需要对生物膜相关活动进行准确的定量和研究。当前的方法通常包括静态生物膜设置（如微孔板中的生物膜测定、珠状生物膜或载玻片上的生物膜）和流体流动生物膜系统（如滴流生物膜反应器、三通道生物膜反应器或管道生物膜反应器）。连续流动系统考虑了流体动力剪切力、营养供应和分散细胞的物理运输的贡献，这些因素定义了大多数自然和工程系统中生物膜发育的栖息地。本方案描述了用于培养PAO1和MR-1模型生物膜的3种流动系统设置的组装，包括各自的定量和观察方法。标准化的流动系统保证了高效且可重复的生物膜实验结果，可根据具体研究项目进一步修改。
