Bone is a complex natural material with a complex hierarchical multiscale organization, crucial to perform its functions. Ultrastructural analysis of bone is crucial for our understanding of cell to cell communication, the healthy or pathological composition of bone tissue, and its three-dimensional (3D) organization. A variety of techniques has been used to analyze bone tissue. This article describes a combined approach of optical, scanning electron, and transmission electron microscopy for the ultrastructural analysis of bone from the nanoscale to the macroscale, as illustrated by two pathological bone tissues. By following a top-down approach to investigate the multiscale organization of pathological bones, quantitative estimates were made in terms of calcium content, nearest neighbor distances of osteocytes, canaliculi diameter, ordering, and D-spacing of the collagen fibrils, and the orientation of intrafibrillar minerals which enable us to observe the fine structural details. We identify and discuss a series of two-dimensional (2D) and 3D imaging techniques that can be used to characterize bone tissue. By doing so we demonstrate that, while 2D imaging techniques provide comparable information from pathological bone tissues, significantly different structural details are observed upon analyzing the pathological bone tissues in 3D. Finally, particular attention is paid to sample preparation for and quantitative processing of data from electron microscopic analysis.

The main aim of this paper is to demonstrate the practical use of nano-indentation and scratch testing in determining mechanical properties of thin coatings. We place our emphasis on how information obtained using both techniques can be combined to give a more complete representation of the properties of a coating-substrate system. Part I of the paper gives an overview of approaches to determine mechanical properties of thin coatings that have been proposed in the literature, and develops them further to be useful tools in the analysis of coatings. This results in methods for measuring the mechanical properties of thin coatings. We particularly emphasise the determination of the elastic modulus, hardness, coating and interfacial fracture toughness and residual stress using indentation and scratch testing. Part II of the paper illustrates the application of these methods to relatively soft coatings of methyltrimethoxysilane (MTMS) filled with colloidal silica or alumina particles on glass. The coatings were prepared using a sol-gel process. We report results of the dependence of the mechanical properties on the filler particle content, illustrating that microstructural changes can also be tracked using these techniques. The effects of the nature and volume fraction of the filler particles are discussed.

The reliability of coatings that are used in industrial applications critically depends on their mechanical properties. Nanoindentation and scratch testing are well-established techniques to measure some of these properties, namely the elastic modulus and hardness of coatings. In this paper, we investigate the possibility of also assessing the coating fracture toughness and the energy of adhesion between the coating and the substrate using indentation and scratch testing. Various existing and new methods are discussed, and they are illustrated by measurements on particle-filled sol-gel coatings on glass. All methods are based on the occurrence of cracking, and they are therefore only applicable to coating systems that act like brittle materials and exhibit cracking during indentation and scratching. The methods for determining the fracture toughness give comparable results, but the values still differ to within about 50%. The values of the adhesion energy obtained from different measurements are consistent, but it remains uncertain to which extent the obtained values are quantitatively correct. The results show that the methods used are promising, but more research is needed to obtain reliable quantitative results.