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Applied and Environmental Microbiology, June 2002, p. 2794-2801, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2794-2801.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Reversible and Irreversible Adhesion of Motile Escherichia coli Cells Analyzed by Total Internal Reflection Aqueous Fluorescence Microscopy

Margot A.-S. Vigeant,1* Roseanne M. Ford,1 Michael Wagner,2 and Lukas K. Tamm2

Department of Chemical Engineering, School of Engineering and Applied Science,1 Department of Molecular Physiology and Biological Physics, School of Medicine, University of Virginia, Charlottesville, Virginia 22904-47412

Received 17 December 2001/ Accepted 20 March 2002

The initial events in bacterial adhesion are often explained as resulting from electrostatic and van der Waals forces between the cell and the surface, as described by DLVO theory (developed by Derjaguin, Landau, Verwey, and Overbeek). Such a theory predicts that negatively charged bacteria will experience greater attraction toward a negatively charged surface as the ionic strength of the medium is increased. In the present study we observed both smooth-swimming and nonmotile Escherichia coli bacteria close to plain, positively, and hydrophobically coated quartz surfaces in high- and low-ionic-strength media by using total internal reflection aqueous fluorescence microscopy. We found that reversibly adhering cells (cells which continue to swim along the surface for extended periods) are too distant from the surface for this behavior to be explained by DLVO-type forces. However, cells which had become immobilized on the surface did seem to be affected by electrostatic interactions. We propose that the "force" holding swimming cells near the surface is actually the result of a hydrodynamic effect, causing the cells to swim at an angle along the glass, and that DLVO-type forces are responsible only for the observed immobilization of irreversibly adhering cells. We explain our observations within the context of a conceptual model in which bacteria that are interacting with the surface may be thought of as occupying one of three compartments: bulk fluid, near-surface bulk, and near-surface constrained. A cell in these compartments feels either no effect of the surface, only the hydrodynamic effect of the surface, or both the hydrodynamic and the physicochemical effects of the surface, respectively.

* Corresponding author. Mailing address: Department of Chemical Engineering, Bucknell University, Lewisburg, PA 17837. Phone: (570) 577-1646. Fax: (570) 577-1141. E-mail: mvigeant{at}bucknell.edu.

Applied and Environmental Microbiology, June 2002, p. 2794-2801, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2794-2801.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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