Lawrence J R, Swerhone G D W, Leppard G G, Araki T, Zhang X, West M M, Hitchcock A P
National Water Research Institute, Saskatoon, Saskatchewan, Canada S7N 3H5.
Appl Environ Microbiol. 2003 Sep;69(9):5543-54. doi: 10.1128/AEM.69.9.5543-5554.2003.
Confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and soft X-ray scanning transmission X-ray microscopy (STXM) were used to map the distribution of macromolecular subcomponents (e.g., polysaccharides, proteins, lipids, and nucleic acids) of biofilm cells and matrix. The biofilms were developed from river water supplemented with methanol, and although they comprised a complex microbial community, the biofilms were dominated by heterotrophic bacteria. TEM provided the highest-resolution structural imaging, CLSM provided detailed compositional information when used in conjunction with molecular probes, and STXM provided compositional mapping of macromolecule distributions without the addition of probes. By examining exactly the same region of a sample with combinations of these techniques (STXM with CLSM and STXM with TEM), we demonstrate that this combination of multimicroscopy analysis can be used to create a detailed correlative map of biofilm structure and composition. We are using these correlative techniques to improve our understanding of the biochemical basis for biofilm organization and to assist studies intended to investigate and optimize biofilms for environmental remediation applications.
共聚焦激光扫描显微镜（CLSM）、透射电子显微镜（TEM）和软X射线扫描透射X射线显微镜（STXM）被用于绘制生物膜细胞和基质中大分子亚组分（如多糖、蛋白质、脂质和核酸）的分布。生物膜由添加了甲醇的河水形成，尽管它们包含复杂的微生物群落，但生物膜以异养细菌为主。TEM提供了最高分辨率的结构成像，CLSM与分子探针结合使用时提供了详细的成分信息，而STXM无需添加探针即可提供大分子分布的成分图谱。通过使用这些技术的组合（STXM与CLSM以及STXM与TEM）精确检查样品的同一区域，我们证明这种多显微镜分析组合可用于创建生物膜结构和组成的详细关联图谱。我们正在使用这些关联技术来增进我们对生物膜组织生化基础的理解，并协助旨在研究和优化用于环境修复应用的生物膜的研究。
