Helmholtz Centre for Environmental Research (UFZ), Magdeburg, Germany.
Environment Canada, Saskatoon, Saskatchewan, Canada.
Trends Microbiol. 2015 Apr;23(4):233-42. doi: 10.1016/j.tim.2014.12.010. Epub 2015 Jan 23.
Confocal laser scanning microscopy has become a standard technique for the investigation of hydrated interfacial microbial communities at the microscale. Multiphoton and spinning-disk microscopes provide new options for in situ imaging. Progress has been made in imaging structural aspects as well as interactions and processes. Advanced fluorescence techniques such as lifetime imaging and correlation spectroscopy are also available. Newly developed target-specific probes allow investigation of new aspects of microbial communities. Several new laser-based techniques are available including nanoscopy and mesoscale techniques. Nanoscopy techniques offer access to unprecedented resolution of hydrated microbiological samples at the scale of fluorescent gene products and macromolecules. Mesoscale approaches are important to address larger features and statistical issues of microbiological samples. This review presents the state of the art in situ biofilm imaging and assesses the pros and cons of laser-based imaging techniques in combination with a variety of sensor types at different scales.
共聚焦激光扫描显微镜已成为研究水合界面微生物群落的标准技术，可以在微观尺度上进行研究。多光子和旋转盘显微镜为原位成像提供了新的选择。在成像结构方面以及相互作用和过程方面都取得了进展。先进的荧光技术，如寿命成像和相关光谱也可应用。新开发的针对特定目标的探针允许研究微生物群落的新方面。几种新的基于激光的技术包括纳米显微镜和中尺度技术。纳米显微镜技术可在荧光基因产物和大分子的尺度上提供对水合微生物样品前所未有的分辨率。中尺度方法对于解决微生物样品的更大特征和统计问题很重要。这篇综述介绍了原位生物膜成像的最新技术，并评估了各种传感器类型在不同尺度下与基于激光的成像技术相结合的优缺点。
