Abstract
To broaden our knowledge on virus structure and function, a profound insight into their mechanical properties is required. Nanoindentation measurements with an atomic force microscope (AFM) are increasingly being performed to probe such material properties. This single-particle approach allows for determining the viral spring constant, their Young’s modulus, as well as the force and deformation at which failure occurs. The experimental procedures for viral nanoindentation experiments are described here in detail, focusing on surface preparation, AFM imaging and nanoindentation, and the subsequent data analysis of the force–distance curves. Whereas AFM can be operated in air and in liquid, the described methods are for probing single viruses in liquid to enable working in a physiologically relevant environment.
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References
Kol, N., Shi, Y., Tsvitov, M., Barlam, D., Shneck, R. Z., Kay, M. S., and Rousso, I. (2007) A stiffness switch in human immunodeficiency virus, Biophys J 92, 1777–83.
Ivanovska, I., Wuite, G., Jonsson, B., and Evilevitch, A. (2007) Internal DNA pressure modifies stability of WT phage, Proc Natl Acad Sci USA 104, 9603–8.
Roos, W. H., Radtke, K., Kniesmeijer, E., Geertsema, H., Sodeik, B., and Wuite, G. J. (2009) Scaffold expulsion and genome packaging trigger stabilization of herpes simplex virus capsids, Proc Natl Acad Sci USA 106, 9673–8.
Ivanovska, I. L., de Pablo, P. J., Ibarra, B., Sgalari, G., MacKintosh, F. C., Carrascosa, J. L., Schmidt, C. F., and Wuite, G. J. L. (2004) Bacteriophage capsids: Tough nanoshells with complex elastic properties, Proc Natl Acad Sci USA 101, 7600–5.
Roos, W. H., Ivanovska, I. L., Evilevitch, A., and Wuite, G. J. L. (2007) Viral capsids: Mechanical characteristics, genome packaging and delivery mechanisms, Cell Mol Life Sci 64, 1484–97.
Roos, W. H., and Wuite, G. J. L. (2009) Nanoindentation studies reveal material properties of viruses, Adv Mater 21, 1187–92.
Hansma, P. K., Cleveland, J. P., Radmacher, M., Walters, D. A., Hillner, P. E., Bezanilla, M., Fritz, M., Vie, D., Hansma, H. G., Prater, C. B., Massie, J., Fukunaga, L., Gurley, J., and Elings, V. (1994) Tapping Mode Atomic-Force Microscopy in Liquids, Appl Phys Lett 64, 1738–40.
Putman, C. A. J., Vanderwerf, K. O., Degrooth, B. G., Vanhulst, N. F., and Greve, J. (1994) Tapping Mode Atomic-Force Microscopy In Liquid, Appl Phys Lett 64, 2454–6.
de Pablo, P. J., Colchero, J., Gomez-Herrero, J., and Baro, A. M. (1998) Jumping mode scanning force microscopy, Appl Phys Lett 73, 3300–2.
Michel, J. P., Ivanovska, I. L., Gibbons, M. M., Klug, W. S., Knobler, C. M., Wuite, G. J. L., and Schmidt, C. F. (2006) Nanoindentation studies of full and empty viral capsids and the effects of capsid protein mutations on elasticity and strength, Proc Natl Acad Sci USA 103, 6184–9.
Carrasco, C., Carreira, A., Schaap, I. A. T., Serena, P. A., Gomez-Herrero, J., Mateu, M. G., and Pablo, P. J. (2006) DNA-mediated anisotropic mechanical reinforcement of a virus, Proc Natl Acad Sci USA 103, 13706–11.
Uetrecht, C., Versluis, C., Watts, N. R., Roos, W. H., Wuite, G. J., Wingfield, P. T., Steven, A. C., and Heck, A. J. (2008) High-resolution mass spectrometry of viral assemblies: Molecular composition and stability of dimorphic hepatitis B virus capsids, Proc Natl Acad Sci USA 105, 9216–20.
Carrasco, C., Castellanos, M., de Pablo, P. J., and Mateu, M. G. (2008) Manipulation of the mechanical properties of a virus by protein engineering, Proc Natl Acad Sci USA 105, 4150–5.
Arkhipov, A., Roos, W. H., Wuite, G. J., and Schulten, K. (2009) Elucidating the mechanism behind irreversible deformation of viral capsids, Biophys J 97, 2061–9.
Sader, J. E., Chon, J. W. M., and Mulvaney, P. (1999) Calibration of rectangular atomic force microscope cantilevers, Rev Sci Instrum 70, 3967–9.
Gibbons, M. M., and Klug, W. S. (2007) Nonlinear finite-element analysis of nanoindentation of viral capsids, Phys Rev E 75, 031901.
Gibbons, M. M., and Klug, W. S. (2008) Influence of nonuniform geometry on nanoindentation of viral capsids, Biophys J 95, 3640–9.
Landau, L. D., and Lifshitz, E. M. (1986) Theory of Elasticity, 3rd ed., Elsevier, Oxford.
Baclayon, M., Wuite, G. J. L., and Roos, W. H. (2010) Imaging and manipulation of single viruses by atomic force microscopy, Soft Matter 6, 5273–85.
Kol, N., Gladnikoff, M., Barlam, D., Shneck, R. Z., Rein, A., and Rousso, I. (2006) Mechanical properties of murine leukemia virus particles: Effect of maturation, Biophys J 91, 767–74.
Klug, W. S., Bruinsma, R. F., Michel, J. P., Knobler, C. M., Ivanovska, I. L., Schmidt, C. F., and Wuite, G. J. L. (2006) Failure of viral shells, Phys Rev Lett 97, 228101.
Liashkovich, I., Hafezi, W., Kuhn, J. E., Oberleithner, H., Kramer, A., and Shahin, V. (2008) Exceptional mechanical and structural stability of HSV-1 unveiled with fluid atomic force microscopy, J Cell Sci 121, 2287–92.
Zhao, Y., Ge, Z. B., and Fang, J. Y. (2008) Elastic modulus of viral nanotubes, Phys Rev E 78, 031914.
Kienberger, F., Zhu, R., Moser, R., Blaas, D., and Hinterdorfer, P. (2004) Monitoring RNA release from human rhinovirus by dynamic force microscopy, J Virol 78, 3203–9.
Xu, X., Carrasco, C., de Pablo, P. J., Gomez-Herrero, J., and Raman, A. (2008) Unmasking imaging forces on soft biological samples in liquids when using dynamic atomic force microscopy: A case study on viral capsids, Biophys J 95, 2520–8.
Butt, H. J., Cappella, B., and Kappl, M. (2005) Force measurements with the atomic force microscope: Technique, interpretation and applications, Surface Science Reports 59, 1–152.
Gibbons, M. M., and Klug, W. S. (2007) Mechanical modeling of viral capsids, J Mater Sci 42, 8995–9004.
Acknowledgments
I would like to thank G.J.L. Wuite and I. Ivanovska for introducing me to the experimental nanoindentation technique and analysis tools. M. Baclayon is thanked for critical reading of the manuscript.
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Roos, W.H. (2011). How to Perform a Nanoindentation Experiment on a Virus. In: Peterman, E., Wuite, G. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 783. Humana Press. https://doi.org/10.1007/978-1-61779-282-3_14
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DOI: https://doi.org/10.1007/978-1-61779-282-3_14
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