Schaudinn C, Carr G, Gorur A, Jaramillo D, Costerton J W, Webster P
Center for Biofilms, USC School of Dentistry, Los Angeles, CA 90089, USA.
J Microsc. 2009 Aug;235(2):124-7. doi: 10.1111/j.1365-2818.2009.03201.x.
Scanning electron microscopy is a useful imaging approach for the visualization of bacterial biofilms in their natural environments including their medical and dental habitats, because it allows for the exploration of large surfaces with excellent resolution of topographic features. Most biofilms in nature, however, are embedded in a thick layer of extracellular matrix that prevents a clear identification of individual bacteria by scanning electron microscopy. The use of confocal laser scanning microscopy on the other hand in combination with fluorescence in situ hybridization enables the visualization of matrix embedded bacteria in multi-layered biofilms. In our study, fluorescence in situ hybridization/confocal laser scanning microscopy and scanning electron microscopy were applied to visualize bacterial biofilm in endodontic root canals. The resulting fluorescence in situ hybridization /confocal laser scanning microscopy and scanning electron microscopy and pictures were subsequently combined into one single image to provide high-resolution information on the location of hidden bacteria. The combined use of scanning electron microscopy and fluorescence in situ hybridization / confocal laser scanning microscopy has the potential to overcome the limits of each single technique.
扫描电子显微镜是一种有用的成像方法，可用于观察细菌生物膜在其自然环境中的情况，包括其在医学和牙科环境中的栖息地，因为它能够以出色的地形特征分辨率探索大面积表面。然而，自然界中的大多数生物膜都嵌入在一层厚厚的细胞外基质中，这使得通过扫描电子显微镜难以清晰识别单个细菌。另一方面，共聚焦激光扫描显微镜与荧光原位杂交相结合，能够观察多层生物膜中嵌入基质的细菌。在我们的研究中，应用荧光原位杂交/共聚焦激光扫描显微镜和扫描电子显微镜来观察根管内的细菌生物膜。随后将所得的荧光原位杂交/共聚焦激光扫描显微镜图像和扫描电子显微镜图像合并为一张单一图像，以提供关于隐藏细菌位置的高分辨率信息。扫描电子显微镜与荧光原位杂交/共聚焦激光扫描显微镜的联合使用有可能克服每种单一技术的局限性。
