AFM Nanoindentation Experiments on Protein Shells:A Protocol

Proteinaceous assemblies are ubiquitous in nature. One important form of these assemblies are protein nanoshells such as viruses; however, a variety of other protein shells exist. To deepen our knowledge on the structure and function of protein shells, 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 us to determine the spring constant, the Young's modulus, as well as the force and deformation at which failure occurs of the nanoshells. The experimental procedures for these mechanical measurements 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 particles in liquid to enable studies in close to physiological environments.