Journal
NATURE PROTOCOLS
Volume 3, Issue 7, Pages 1132-1138Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2008.101
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Funding
- National Foundation for Scientific Research (FNRS)
- Universite Catholique de Louvain (Fonds Speciaux de Recherche)
- Region wallonne
- Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme)
- Research Department of the Communaute francaise de Belgique (Concerted Research Action)
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Over the past years, atomic force microscopy (AFM) has emerged as a powerful tool for imaging the surface of microbial cells with nanometer resolution, and under physiological conditions. Moreover, chemical force microscopy (CFM) and single-molecule force spectroscopy have enabled researchers to map chemical groups and receptors on cell surfaces, providing valuable insight into their structure-function relationships. Here, we present protocols for analyzing spores of the pathogen Aspergillus fumigatus using real-time AFM imaging and CFM. We emphasize the use of porous polymer membranes for immobilizing single live cells, and the modification of gold-coated tips with alkanethiols for CFM measurements. We also discuss recording conditions and data interpretation, and provide recommendations for reliable experiments. For well-trained AFM users, the entire protocol can be completed in 2-3 d.
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