Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

Author:

Lawrence J. R.1,Swerhone G. D. W.1,Leppard G. G.2,Araki T.3,Zhang X.3,West M. M.4,Hitchcock A. P.3

Affiliation:

1. National Water Research Institute, Saskatoon, Saskatchewan, Canada S7N 3H5

2. NWRI, Burlington, Ontario, Canada L7R 4A6

3. Brockhouse Institute for Materials Research, McMaster University, Hamilton, Ontario, Canada L8S 4M1

4. Electron Microscopy Facility, Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada L8N 3Z5

Abstract

ABSTRACT 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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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