Jani Sneha
Molecular Foundry, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Methods Mol Biol. 2018;1729:61-69. doi: 10.1007/978-1-4939-7577-8_6.
Bacterial chemotaxis in response to continuous chemical gradients has been extensively studied at the individual cell and population levels using a variety of well-established in vitro methods (Englert et al., Microfluidic techniques for the analysis of bacterial chemotaxis. Methods Mol Biol 571:1-23, 2009). In nature, bacteria are surrounded by heterogeneous chemical gradients; hence, it is essential to understand chemotaxis behavior under such conditions. Here, we describe a setup that allows visualization of the chemotaxis response of motile cells to the complex microenvironment of a biofilm maintained under static conditions. The biofilm is separated from the motile cells by a semi-permeable membrane. Cells swimming toward the biofilm are captured on the membrane and imaged using confocal laser scanning microscopy (CLSM). Chemotaxis toward specific molecules produced by the biofilm, such as autoinducer-2 (AI-2), can be studied using this setup. This system can be adapted to study chemotaxis toward poly-species biofilms, or even mammalian cells.
利用各种成熟的体外方法，在单个细胞和群体水平上对细菌响应连续化学梯度的趋化性进行了广泛研究（恩格勒特等人，用于分析细菌趋化性的微流控技术。《分子生物学方法》571:1-23，2009）。在自然界中，细菌被异质化学梯度所包围；因此，了解在这种条件下的趋化行为至关重要。在这里，我们描述了一种装置，该装置可以观察运动性细胞对在静态条件下维持的生物膜复杂微环境的趋化反应。生物膜通过半透膜与运动性细胞分离。向生物膜游动的细胞被捕获在膜上，并使用共聚焦激光扫描显微镜（CLSM）成像。利用该装置可以研究对生物膜产生的特定分子（如自诱导物-2，AI-2）的趋化性。该系统可用于研究对多物种生物膜甚至哺乳动物细胞的趋化性。
