Pseudomonas aeruginosa and Saccharomyces cerevisiae Biofilm in Flow Cells
Pseudomonas aeruginosa and Saccharomyces cerevisiae Biofilm in Flow Cells
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DOI：
10.3791/2383
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发表时间：
2011-01-01
影响因子：
1.2
通讯作者：
Regenberg, Birgitte
中科院分区：
文献类型：
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作者：
Nielsen, Martin Weiss;Sternberg, Claus;Regenberg, Birgitte
许多微生物细胞能够形成固着微生物群落，被定义为生物膜，与自由生活的微生物相比，其生理和病理特性发生了改变。自然界中的生物膜通常难以研究并且存在于定义不明确的条件下(1)。使用透明基质，可以装置一个可以以非破坏性方式实时检查简单生物膜的系统：在这里，我们演示了流动池模型系统的组装和操作，用于微生物生物膜的体外 3D 研究，在明确的条件下产生高重现性 (2,3)。该系统由充当生物膜生长室的流动池组成。通过蠕动泵从培养基烧瓶向流动池提供营养物和氧气，用过的培养基收集在废物容器中。流动系统的这种结构允许连续供应营养物和施用大肠杆菌。 g。抗生素对流动室中生长的细胞的干扰最小。此外，流动池内的流动条件允许研究暴露于剪切应力的生物膜。气泡捕获装置限制管道中的气泡，否则可能会破坏流动池中的生物膜结构。流动池系统与共焦激光扫描显微镜 (CLSM) 兼容，从而可以提供有关正在形成的微生物生物膜的高度详细的 3D 信息。生物膜中的细胞可以用与 CLSM 分析兼容的荧光探针或蛋白质进行标记。这使得在线可视化成为可能，并允许研究正在发育的生物膜中的生态位。微生物之间的相互关系、抗菌剂的研究或特定基因的表达，是可以在流动池系统中研究的许多实验设置之一。
Many microbial cells have the ability to form sessile microbial communities defined as biofilms that have altered physiological and pathological properties compared to free living microorganisms. Biofilms in nature are often difficult to investigate and reside under poorly defined conditions(1). Using a transparent substratum it is possible to device a system where simple biofilms can be examined in a non-destructive way in real-time: here we demonstrate the assembly and operation of a flow cell model system, for in vitro 3D studies of microbial biofilms generating high reproducibility under well-defined conditions(2,3).The system consists of a flow cell that serves as growth chamber for the biofilm. The flow cell is supplied with nutrients and oxygen from a medium flask via a peristaltic pump and spent medium is collected in a waste container. This construction of the flow system allows a continuous supply of nutrients and administration of e. g. antibiotics with minimal disturbance of the cells grown in the flow chamber. Moreover, the flow conditions within the flow cell allow studies of biofilm exposed to shear stress. A bubble trapping device confines air bubbles from the tubing which otherwise could disrupt the biofilm structure in the flow cell.The flow cell system is compatible with Confocal Laser Scanning Microscopy (CLSM) and can thereby provide highly detailed 3D information about developing microbial biofilms. Cells in the biofilm can be labeled with fluorescent probes or proteins compatible with CLSM analysis. This enables online visualization and allows investigation of niches in the developing biofilm. Microbial interrelationship, investigation of antimicrobial agents or the expression of specific genes, are of the many experimental setups that can be investigated in the flow cell system.