Quantification and localization of the liquid zone of partially remelted M2 tool steel using X-ray microtomography and scanning electron microscopy

The authors warmly thank Luc Morhain and Marc Wary (Arts et Métiers ParisTech CER Metz) for their technical support.Thixoforming of steels poses challenges due to the high temperatures involved and the lack of understanding of thermomechanical behavior. The volume fractions of the liquid and solid phases in the semi-solid state are the most important parameters for such a form-ing process, as they affect the viscosity and hence the flow behavior of the material. Two-dimensional observations might not always be sufficient, as the size distribution and the connectivity of phases cannot be obtained from associated measurements, which can only be determined by three-dimensional (3-D) investigation. This paper presents the first application of high-energy X-ray microtomography to the microstructure of steel in the semi-solid state. The microstructure of M2 high-speed tool steel was studied in both as-received and heated-and-quenched states. From the reconstructed images, 3-D information could be obtained and was compared with scanning elec-tron microscopy and energy dispersive spectrometry observations. The volume fraction and the location of liquid phase in the semi-solid state were determined in particular, and the continuous solid skeleton was investigated

3D Multicolor Super-Resolution Imaging Offers Improved Accuracy in Neuron Tracing

The connectivity among neurons holds the key to understanding brain function. Mapping neural connectivity in brain circuits requires imaging techniques with high spatial resolution to facilitate neuron tracing and high molecular specificity to mark different cellular and molecular populations. Here, we tested a three-dimensional (3D), multicolor super-resolution imaging method, stochastic optical reconstruction microscopy (STORM), for tracing neural connectivity using cultured hippocampal neurons obtained from wild-type neonatal rat embryos as a model system. Using a membrane specific labeling approach that improves labeling density compared to cytoplasmic labeling, we imaged neural processes at 44 nm 2D and 116 nm 3D resolution as determined by considering both the localization precision of the fluorescent probes and the Nyquist criterion based on label density. Comparison with confocal images showed that, with the currently achieved resolution, we could distinguish and trace substantially more neuronal processes in the super-resolution images. The accuracy of tracing was further improved by using multicolor super-resolution imaging. The resolution obtained here was largely limited by the label density and not by the localization precision of the fluorescent probes. Therefore, higher image resolution, and thus higher tracing accuracy, can in principle be achieved by further improving the label density

Quantification of sub-resolution porosity in carbonate rocks by applying high-salinity contrast brine using X-ray microtomography differential imaging

Characterisation of the pore space in carbonate reservoirs and aquifers is of utmost importance in a number of applications such as enhanced oil recovery, geological carbon storage and contaminant transport. We present a new experimental methodology that uses high-salinity contrast brine and differential imaging acquired by X-ray tomography to non-invasively obtain three-dimensional spatially resolved information on porosity and connectivity of two rock samples, Portland and Estaillades limestones, including sub-resolution micro-porosity. We demonstrate that by injecting 30 wt% KI brine solution, a sufficiently high phase contrast can be achieved allowing accurate three-phase segmentation based on differential imaging. This results in spatially resolved maps of the solid grain phase, sub-resolution micro-pores within the grains, and macro-pores. The total porosity values from the three-phase segmentation for two carbonate rock samples are shown to be in good agreement with Helium porosity measurements. Furthermore, our flow-based method allows for an accurate estimate of pore connectivity and a distribution of porosity within the sub-resolution pores

Image informatics strategies for deciphering neuronal network connectivity

Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphologi- cal plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular com- munication and the associated calcium-bursting behaviour. In vitro cultured neu- ronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies