Abstract
Functionalization of AFM probes with biomolecules or microorganisms allows for a better understanding of the interaction mechanisms driving microbial adhesion. Here we describe the most commonly used protocols to graft molecules and bacteria to AFM cantilevers. The bioprobes obtained that way enable to measure forces down to the single-cell and single-molecule levels.
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References
Beaussart A, Pechoux C, Trieu-Cuot P et al (2014) Molecular mapping of the cell wall polysaccharides of the human pathogen Streptococcus agalactiae. Nanoscale 6:14820–14827
Dover RS, Bitler A, Shimoni E et al (2015) Multiparametric AFM reveals turgor-responsive net-like peptidoglycan architecture in live streptococci. Nat Commun 6:7193
Hayhurst EJ, Kailas L, Hobbs JK et al (2008) Cell wall peptidoglycan architecture in Bacillus subtilis. Proc Natl Acad Sci U S A 105:14603–14608
Turner RD, Ratcliffe EC, Wheeler R et al (2010) Peptidoglycan architecture can specify division planes in Staphylococcus aureus. Nat Commun 1:1
Andre G, Kulakauskas S, Chapot-Chartier MP et al (2010) Imaging the nanoscale organization of peptidoglycan in living Lactococcus lactis cells. Nat Commun 1:1
El-Kirat-Chatel S, Dufrene YF (2012) Nanoscale imaging of the Candida - macrophage interaction using correlated fluorescence-atomic force microscopy. ACS Nano 6:10792–10799
Labernadie A, Thibault C, Vieu C et al (2010) Dynamics of podosome stiffness revealed by atomic force microscopy. Proc Natl Acad Sci U S A 107:21016–21021
Dupres V, Alsteens D, Pauwels K et al (2009) In vivo imaging of S-layer Nanoarrays on Corynebacterium glutamicum. Langmuir 25:9653–9655
Alsteens D, Aimanianda V, Hegde P et al (2013) Unraveling the nanoscale surface properties of chitin synthase mutants of Aspergillus fumigatus and their biological implications. Biophys J 105:320–327
Bayry J, Beaussart A, Dufrene YF et al (2014) Surface structure characterization of Aspergillus fumigatus conidia mutated in the melanin synthesis pathway and their human cellular immune response. Infect Immun 82:3141–3153
Chtcheglova LA, Hinterdorfer P (2011) Simultaneous topography and recognition imaging on endothelial cells. J Mol Recognit 24:788–794
El-Kirat-Chatel S, Boyd CD, O’Toole GA et al (2014) Single-molecule analysis of Pseudomonas fluorescens footprints. ACS Nano 8:1690–1698
Heinisch JJ, Lipke PN, Beaussart A et al (2012) Atomic force microscopy – looking at mechanosensors on the cell surface. J Cell Sci 125:4189–4195
Hinterdorfer P, Dufrene YF (2006) Detection and localization of single molecular recognition events using atomic force microscopy. Nat Methods 3:347–355
Muller DJ, Dufrene YF (2011) Atomic force microscopy: a nanoscopic window on the cell surface. Trends Cell Biol 21:461–469
Alsteens D, Garcia MC, Lipke PN et al (2010) Force-induced formation and propagation of adhesion nanodomains in living fungal cells. Proc Natl Acad Sci U S A 107:20744–20749
Beaussart A, Alsteens D, El-Kirat-Chatel S et al (2012) Single-molecule imaging and functional analysis of Als Adhesins and Mannans during Candida albicans morphogenesis. ACS Nano 6:10950–10964
Heinisch JJ, Dupres V, Wilk S et al (2010) Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1. PLoS One 5:e11104
Alsteens D, Dupres V, Klotz SA et al (2009) Unfolding individual Als5p adhesion proteins on live cells. ACS Nano 3:1677–1682
El-Kirat-Chatel S, Beaussart A, Boyd CD et al (2014) Single-cell and single-molecule analysis deciphers the localization, adhesion, and mechanics of the biofilm Adhesin LapA. ACS Chem Biol 9:485–494
Francius G, Alsteens D, Dupres V et al (2009) Stretching polysaccharides on live cells using single molecule force spectroscopy. Nat Protoc 4:939–946
Marszalek PE, Oberhauser AF, Pang YP et al (1998) Polysaccharide elasticity governed by chair-boat transitions of the glucopyranose ring. Nature 396:661–664
Rief M, Oesterhelt F, Heymann B et al (1997) Single molecule force spectroscopy on polysaccharides by atomic force microscopy. Science 275:1295–1297
Beaussart A, Ngo TC, Derclaye S et al (2014) Chemical force microscopy of stimuli-responsive adhesive copolymers. Nanoscale 6:565–571
Alsteens D, Dague E, Rouxhet PG et al (2007) Direct measurement of hydrophobic forces on cell surfaces using AFM. Langmuir 23:11977–11979
Alsteens D, Dupres V, Yunus S et al (2012) High-resolution imaging of chemical and biological sites on living cells using peak force tapping atomic force microscopy. Langmuir 28:16738–16744
Dague E, Alsteens D, Latge JP et al (2007) Chemical force microscopy of single live cells. Nano Lett 7:3026–3030
Dufrene YF (2008) Atomic force microscopy and chemical force microscopy of microbial cells. Nat Protoc 3:1132–1138
Beaussart A, El-Kirat-Chatel S, Sullan RMA et al (2014) Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy. Nat Protoc 9:1049–1055
Alsteens D, Beaussart A, Derclaye S et al (2013) Single-cell force spectroscopy of Als-mediated fungal adhesion. Anal Methods 5:3657–3662
Beaussart A, El-Kirat-Chatel S, Herman P et al (2013) Single-cell force spectroscopy of probiotic Bacteria. Biophys J 104:1886–1892
El-Kirat-Chatel S, Dufrene YF (2016) Nanoscale adhesion forces between the fungal pathogen Candida albicans and macrophages. Nanoscale Horiz 1:69–74
Helenius J, Heisenberg CP, Gaub HE et al (2008) Single-cell force spectroscopy. J Cell Sci 121:1785–1791
Jauvert E, Dague E, Severac M et al (2012) Probing single molecule interactions by AFM using bio-functionalized dendritips. Sens Actuators B Chem 168:436–441
Meister A, Gabi M, Behr P et al (2009) FluidFM: combining atomic force microscopy and nanofluidics in a universal liquid delivery system for single cell applications and beyond. Nano Lett 9:2501–2507
Ebner A, Wildling L, Kamruzzahan ASM et al (2007) A new, simple method for linking of antibodies to atomic force microscopy tips. Bioconjug Chem 18:1176–1184
Wildling L, Unterauer B, Zhu R et al (2011) Linking of sensor molecules with amino groups to amino-functionalized AFM tips. Bioconjug Chem 22:1239–1248
Dupres V, Alsteens D, Wilk S et al (2009) The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo. Nat Chem Biol 5:857–862
Beaussart A, Abellan-Flos M, El-Kirat-Chatel S et al (2016) Force nanoscopy as a versatile platform for quantifying the activity of antiadhesion compounds targeting bacterial pathogens. Nano Lett 16:1299–1307
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El-Kirat-Chatel, S., Beaussart, A. (2018). Probing Bacterial Adhesion at the Single-Molecule and Single-Cell Levels by AFM-Based Force Spectroscopy. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_24
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DOI: https://doi.org/10.1007/978-1-4939-8591-3_24
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