Imaging Bacterial Cells and Biofilms Adhering to Hydrophobic Organic Compound–Water Interfaces
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Assimilation of hydrophobic organic compound (HOC) entails frequently the formation of biofilm at the HOC–water interface. Knowledge on the behavior of cells at the oil–water interface and within the structured biofilm is therefore important to understand the degradation of the HOC in ecosystems. The adhesion and biofilm formation on oil–water interface are best documented by microscopic observations. In this chapter we thus describe two methods for observation of bacterial cells and biofilms growing at the HOC–water interface. The first method uses CLSM (confocal laser scanning microscopy) to obtain in situ images of biofilm developing on thin paraffin strip which offers a flat transparent surface allowing imaging directly through the bottom of the culture dish without sampling. Alternatively, the biofilm can be grown on a paraffin strip deposited on a glass microscope slide and then imaged from the top when high resolution is needed. The second method addresses the problem of the ultrastructure of biofilm developing on HOC. It enables to obtain by TEM (transmission electron microscopy) images of cross sections of biofilms with identification of the side in contact with the HOC.
Keywords:Adhesion Biofilm CLSM Hydrocarbons Lipids Oleolytic bacteria TEM
We thank the MIMA2 platform (www.jouy.inra.fr/mima2) for its expertise and access to microscopy equipments and Michael Trichet (Institut de Biologie Paris-Seine, Université P. et M. Curie) for his advices and comments for TEM. We gratefully acknowledge the French National Research Agency, project AD’HOC ANR-11-BSV7-0002, for financial support.
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