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Applied and Environmental Microbiology, September 2008, p. 5457-5465, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.02075-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Use of Atomic Force Microscopy and Transmission Electron Microscopy for Correlative Studies of Bacterial Capsules{triangledown} ,{dagger}

Oleg Stukalov,1,3 Anton Korenevsky,2,3 Terry J. Beveridge,2,3 and John R. Dutcher1,3*

Department of Physics,1 Department of Molecular and Cellular Biology,2 Advanced Foods and Materials Network, Networks of Centres of Excellence, University of Guelph, Guelph, Ontario, Canada N1G 2W13

Received 11 September 2007/ Accepted 18 June 2008

Bacteria can possess an outermost assembly of polysaccharide molecules, a capsule, which is attached to their cell wall. We have used two complementary, high-resolution microscopy techniques, atomic force microscopy (AFM) and transmission electron microscopy (TEM), to study bacterial capsules of four different gram-negative bacterial strains: Escherichia coli K30, Pseudomonas aeruginosa FRD1, Shewanella oneidensis MR-4, and Geobacter sulfurreducens PCA. TEM analysis of bacterial cells using different preparative techniques (whole-cell mounts, conventional embeddings, and freeze-substitution) revealed capsules for some but not all of the strains. In contrast, the use of AFM allowed the unambiguous identification of the presence of capsules on all strains used in the present study, including those that were shown by TEM to be not encapsulated. In addition, the use of AFM phase imaging allowed the visualization of the bacterial cell within the capsule, with a depth sensitivity that decreased with increasing tapping frequency.

* Corresponding author. Mailing address: Department of Physics, University of Guelph, 50 Stone Rd. E, Guelph, Ontario, Canada N1G 2W1. Phone: (519) 824-4120. Fax: (519) 836-9967. E-mail: dutcher{at}physics.uoguelph.ca

{triangledown} Published ahead of print on 7 July 2008.

{dagger} This work is dedicated to T.J.B., our good friend and superb colleague, who passed away during the preparation of the manuscript.

Applied and Environmental Microbiology, September 2008, p. 5457-5465, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.02075-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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