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Characterization of Pseudomonas fluorescens Biofilm

Blankemeier, Andrew R.
http://rave.ohiolink.edu/etdc/view?acc_num=osu1307731184

Abstract Details

2011, Master of Science, Ohio State University, Materials Science and Engineering.
Biofilms are communities of bacteria that live on substrates. In many cases, biofilms have a detrimental effect on the surfaces to which they adhere. For example, biofilms negatively impact dental enamel, metals, and polymers. One of many challenges in biofilm research is analyzing the interface between a biofilm and its substrate, to determine how the biofilm adheres to, and affects, the substrate. This analysis can be challenging due to the limitations of methods such as SEM, confocal microscopy and ultramicrotomy. Indeed, SEM can provide topographical information of the sample’s surface but is fundamentally a two dimensional technique. Confocal microscopy is a useful tool for characterizing large areas of the biofilm but lacks the spatial resolution that TEM provides. Traditionally, ultramicrotomy has been the preferred method of sample preparation for TEM analysis of biological systems despite the lack of site-specificity and compromising effect on structure from mechanical sectioning with a knife. Recently, dual-beam focused ion beam (DB-FIB) technology has revolutionized the field of metals, ceramics, semiconductor materials, and TEM sample preparation; however this technique has not been widely applied to soft material characterization largely due to rapid degradation of these materials under ion beam irradiation. In this study, we have developed novel processes to reduce ion damage to the biofilm during milling, and successfully used DB-FIB to analyze Pseudomonas fluorescens biofilms and the interface between the biofilm and its substrate, polyurethane paint. An FEI Helios Nanolab 600 DB-FIB was used to create serial section data sets for three-dimensional reconstruction and also to excise thin foils for (Scanning) Transmission Electron Microscopy ((S)TEM) and Electron Energy Loss Spectroscopy (EELS). Serial sectioning of the biofilm provided novel structural information about surface adherence mechanisms of the bacteria. (S)TEM imaging and EELS characterization was applied to investigate compositional differences across the interface as well as view the internal structures (or inclusions) of the bacteria.
Hamish Fraser (Advisor)
William Clark (Committee Member)
61 p.
Biology; Engineering; Microbiology
Characterization; Biofilm; FIB; TEM; SEM;

Recommended Citations

Citations

  • Blankemeier, A. R. (2011). Characterization of Pseudomonas fluorescens Biofilm [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1307731184

    APA Style (7th edition)

  • Blankemeier, Andrew. Characterization of Pseudomonas fluorescens Biofilm. 2011. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1307731184.

    MLA Style (8th edition)

  • Blankemeier, Andrew. "Characterization of Pseudomonas fluorescens Biofilm." Master's thesis, Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1307731184

    Chicago Manual of Style (17th edition)

osu1307731184
443
© 2011, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.