The use of microscopy and three-dimensional visualization to evaluate the structure of microbial biofilms cultivated in the Calgary Biofilm Device
The use of microscopy and three-dimensional visualization to evaluate the structure of microbial biofilms cultivated in the Calgary Biofilm Device
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DOI：
10.1251/bpo127
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发表时间：
2006-12-19
影响因子：
6.4
通讯作者：
Turner, Raymond J.
中科院分区：
文献类型：
--
作者：
Harrison, Joe J.;Ceri, Howard;Turner, Raymond J.
微生物通常生活在多细胞的固体表面附着的集合体中，称为生物膜。这些微生物群落具有结构特征，有助于群体异质性，从而促进细胞功能的产生。因此，生物膜结构的三维（3D）特征对于理解这些微生物系统的生理和生态是重要的。本文详细介绍了几种协议的扫描电子显微镜和共聚焦激光扫描显微镜（CLSM）的聚苯乙烯钉在卡尔加里生物膜装置（CBD）上生长的生物膜。此外，描述了使用虚拟现实工具amira（TM）对CLSM数据栈进行图像处理以创建生物膜微生物的表面和/或体积渲染的3D可视化的过程。显微镜与CBD中的微生物培养相结合-一种设计用于高通量敏感性测试的装置-允许在多变量生长和暴露条件下对生物膜进行结构-功能分析。
Microbes frequently live within multicellular, solid surface-attached assemblages termed biofilms. These microbial communities have architectural features that contribute to population heterogeneity and consequently to emergent cell functions. Therefore, three-dimensional (3D) features of biofilm structure are important for understanding the physiology and ecology of these microbial systems. This paper details several protocols for scanning electron microscopy and confocal laser scanning microscopy (CLSM) of biofilms grown on polystyrene pegs in the Calgary Biofilm Device (CBD). Furthermore, a procedure is described for image processing of CLSM data stacks using amira (TM), a virtual reality tool, to create surface and/or volume rendered 3D visualizations of biofilm microorganisms. The combination of microscopy with microbial cultivation in the CBD - an apparatus that was designed for high-throughput susceptibility testing - allows for structure-function analysis of biofilms under multivariate growth and exposure conditions.