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Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy

Nature Protocols volume 9, pages 1049–1055 (2014)Cite this article

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Abstract

During the past decades, several methods (e.g., electron microscopy, flow chamber experiments, surface chemical analysis, surface charge and surface hydrophobicity measurements) have been developed to investigate the mechanisms controlling the adhesion of microbial cells to other cells and to various other substrates. However, none of the traditional approaches are capable of looking at adhesion forces at the single-cell level. In recent years, atomic force microscopy (AFM) has been instrumental in measuring the forces driving microbial adhesion on a single-cell basis. The method, known as single-cell force spectroscopy (SCFS), consists of immobilizing a single living cell on an AFM cantilever and measuring the interaction forces between the cellular probe and a solid substrate or another cell. Here we present SCFS protocols that we have developed for quantifying the cell adhesion forces of medically important microbes. Although we focus mainly on the probiotic bacterium Lactobacillus plantarum, we also show that our procedures are applicable to pathogens, such as the bacterium Staphylococcus epidermidis and the yeast Candida albicans. For well-trained microscopists, the entire protocol can be mastered in 1 week.

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Figure 1: Principles of SCFS.
Figure 2: Measurement of the specific (glycopolymer) and nonspecific (hydrophobic) interactions of probiotic bacteria.
Figure 3: Quantification of the specific forces engaged in bacterial-fungal interactions.

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Acknowledgements

Work at the Université Catholique de Louvain was supported by the National Foundation for Scientific Research (FNRS), the Université Catholique de Louvain (Fondation Louvain-Prix De Merre), the Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme) and the Research Department of the Communauté Française de Belgique (Concerted Research Action). Y.F.D. and D.A. are Research Director and Postdoctoral Researcher of the Fonds de la Recherche Scientifique (FRS)-FNRS, respectively.

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Authors and Affiliations

  1. Université Catholique de Louvain, Institute of Life Sciences, Louvain-la-Neuve, Belgium

    Audrey Beaussart, Sofiane El-Kirat-Chatel, Ruby May A Sullan, David Alsteens, Philippe Herman, Sylvie Derclaye & Yves F Dufrêne

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  1. Audrey Beaussart
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Contributions

A.B., S.E.-K.-C., R.M.A.S., D.A., P.H., S.D. and Y.F.D. designed the research; A.B., S.E.-K.-C., R.M.A.S., D.A., P.H. and S.D. performed the research; A.B., S.E.-K.-C., R.M.A.S., D.A., P.H., S.D. and Y.F.D. analyzed the data and wrote the paper.

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Correspondence to Yves F Dufrêne.

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The authors declare no competing financial interests.

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Beaussart, A., El-Kirat-Chatel, S., Sullan, R. et al. Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy. Nat Protoc 9, 1049–1055 (2014). https://doi.org/10.1038/nprot.2014.066

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