Identification of ontogenetic age classes plays an important role in the fields of zoology, palaeontology and archaeology, where accurate age classifications of (sub)fossil remains are a crucial component for the reconstruction of past life. Textural ageing—the identification of age-related bone surface textures—provides a size-independent method for age assessment of vertebrate material. However, most of the work so far is limited to qualitative results. While qualitative approaches provide helpful insights on textural ageing patterns, they are heavily subject to observer bias and fall short of quantitative data relevant for detailed statistical analyses and cross-comparisons. Here, we present a pilot study on the application of 3D surface digital microscopy to quantify bone surface textures on the long bones of the grey heron (Ardea cinerea) and the Canada goose (Branta canadensis) using internationally verified roughness parameters. Using a standardised measuring protocol, computed roughness values show a strong correlation with qualitative descriptions of textural patterns. Overall, higher roughness values correspond to increased numbers of grooves and pits and vice versa. Most of the roughness parameters allowed distinguishing between different ontogenetic classes and closely followed the typical sigmoidal animal growth curve. Our results show that bone texture quantification is a feasible approach to identifying ontogenetic age classes.

Electron beam lithography (EBL) requires conducting substrates to ensure pattern fidelity. However, there is an increasing interest in performing EBL on less well-conducting surfaces or even insulators, usually resulting in seriously distorted pattern formation. To understand the underlying charging phenomena, the authors use Monte Carlo simulations that include models for substrate charging, electron beam-induced current, and electric breakdown. Simulations of electron beam exposure of glass wafers are presented, exposing regular patterns which become distorted due to charge-induced beam deflection. The resulting displacements within the patterns are mapped and compared to experimental displacement maps obtained from patterns in PMMA resist on glass substrates. Displacements up to several hundreds of nanometers were observed at a primary beam energy of 50 keV. Also, various scan strategies were used to write the patterns, in the simulations as well as the experiments, revealing their strong effect on pattern distortion, in shape and in magnitude. A qualitative, in some cases even quantitative, good agreement was found between the simulations and the experiments, providing enough confidence in Monte Carlo simulations to predict charge-induced pattern displacement and shape distortion and to find smart scan strategies to minimize the effects of charging.

Recent research evidences the strong modulatory role of controlled submicron and nanoscale topographies on stem cells fate. To harness these physical surface cues for clinical applications, fabrication of nano- and submicron patterns on clinically relevant biomaterials is greatly needed. In this study, an electron beam lithography method for direct patterning (i.e., no use of masters/imprinting steps) of titanium in the submicron range was developed. The process required the use of an etch mask consisting of a double layer of SiO₂ and Al, and the positive AR P-6200.04 electron beam resist. An optimum electron beam dose of 288 μC/cm² was established for writing the desired patterns. The transfer of the patterns into the titanium substrates was achieved by three different steps: inductively coupled plasma etching of the mask in BCl₃/Cl₂ followed by reactive ion etching of titanium in SF₆/CHF₃/O₂ and a final wet etch of mask residue. Highly ordered arrays of titanium pits with submicron diameters were produced with high reproducibility. This method provides great versatility in pattern design, direct transfer into titanium and increased control of titanium pattern features at submicron to nanoscale enabling clinically relevant and systematic studies on pattern-induced cellular responses.