Atomic force microscopy and chemical force microscopy of microbial cells.
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.
Dufrêne
Yves F
YF
Unité de Chimie des Interfaces, Université Catholique de Louvain, Croix du Sud 2/18, B-1348 Louvain-la-Neuve, Belgium. Yves.Dufrene@uclouvain.be
D001232:Aspergillus fumigatus / Q000648:ultrastructure*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force / Q000379:methods*; D013172:Spores, Fungal / Q000648:ultrastructure*
D008567:Membranes, Artificial