Abstract
Microbially induced corrosion (MIC) is an emerging topic that has huge environmental impacts, such as long-term evaluation of microbial interactions with radioactive waste glass, environmental cleanup and disposal of radioactive material, and weathering effects of microbes. Time-of-flight secondary ion mass spectrometry (ToF-SIMS), a powerful mass spectral imaging technique with high surface sensitivity, mass resolution, and mass accuracy, can be used to study biofilm effects on different substrates. Understanding how to prepare biofilms on MIC susceptible substrates is critical for proper analysis via ToF-SIMS. We present here a step-by-step protocol for preparing bacterial biofilms for ToF-SIMS analysis, comparing three biofilm preparation techniques: no desalination, centrifugal spinning (CS), and water submersion (WS). Comparisons of two desalinating methods, CS and WS, show a decrease in the media peaks up to 99% using CS and 55% using WS, respectively. Proper desalination methods also can increase biological signals by over four times for fatty acids using WS, for example. ToF-SIMS spectral results show chemical compositional changes of the glass exposed in a Paenibacillus polymyxa SCE2 biofilm, indicating its capability to probe microbiologically induced corrosion of solid surfaces. This represents the proper desalination technique to use without significantly altering biofilm structure and substrate for ToF-SIMS analysis. ToF-SIMS spectral results showed chemical compositional changes of the glass exposed by a Paenibacillus bacterial biofilm over 3-month inoculation. Possible MIC products include various phosphate phase molecules not observed in any control samples with the highest percent increases when experimental samples were compared with biofilm control samples.
Original language | English |
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Article number | 051004 |
Journal | Biointerphases |
Volume | 19 |
Issue number | 5 |
DOIs | |
State | Published - Sep 1 2024 |
Funding
Research efforts were supported by the strategic Laboratory Directed Research and Development (LDRD) of the Physical Sciences Directorate of the Oak Ridge National Laboratory (ORNL). The authors thank Jiyoung Son, Zihua Zhu, and Jeffery Dhas for their support in sample preparation and analysis. The authors thank Jose Marcial for synthesis of the glass and William Chrisler for collection of the CLSM data. The authors also thank James Neeway and Carolyn Pearce for access to glass samples of the Hillfort Glass project. The authors are indebted to Haluk Beyenal and Janet Jansson for their contributions of bacterial strains. A portion of the research was performed using EMSL (No. grid.436923.9), a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research under the general EMSL user Proposal Nos. 60770 and 61024 at the Pacific Northwest National Laboratory (PNNL). ORNL is managed by UT-Battelle, LLC, for the U.S. DOE under Contract No. DE-AC05-00OR22725. PNNL is operated by Battelle for the DOE under Contract No. DE-AC05-76RL01830. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. The DOE will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan (http://energy.gov/downloads/doe-public-access-plan). Gabriel Parker was funded by the Department of Energy SCGSR fellowship program at Pacific Northwest National Laboratory (PNNL) and by the GRO internship program at the Oak Ridge National Laboratory (ORNL). Xiao-Ying Yu is indebted to the support of the strategic Laboratory Directed Research and Development (LDRD) project of the Physical Sciences Directorate of ORNL.
Funders | Funder number |
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United States Government | |
DOE Public Access Plan | |
Pacific Northwest National Laboratory | |
Oak Ridge National Laboratory | |
Laboratory Directed Research and Development | |
Office of Science | |
Biological and Environmental Research | 61024, 60770 |
U.S. Department of Energy | DE-AC05-00OR22725, DE-AC05-76RL01830 |