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Abstract: Integrating physiology with structural insights of the same neuronal circuit provides a unique approach to understanding how the mammalian brain computes information. However, combining the techniques that provide both streams of data represents an experimental challenge. When studying glomerular column circuits in the mouse olfactory bulb, this approach involves e.g., recording the neuronal activity with in vivo 2-photon (2P) calcium imaging, retrieving the circuit structure with synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT) and/or serial block-face scanning electron microscopy (SBEM) and correlating these datasets. Sample preparation and dataset correlation are two key bottlenecks in this correlative workflow. Here, we first quantify the occurrence of different artefacts when staining tissue slices with heavy metals to generate X-ray or electron contrast. We report improvements in the staining procedure, ultimately achieving perfect staining in ∼67% of the 0.6 mm thick olfactory bulb slices that were previously imaged in vivo with 2P. Secondly, we characterise the accuracy of the spatial correlation between functional and structural datasets. We demonstrate that direct, single-cell precise correlation between in vivo 2P and SXRT tissue volumes is possible and as reliable as correlating between 2P and SBEM. Altogether, these results pave the way for experiments that require retrieving physiology, circuit structure and synaptic signatures in targeted regions. These correlative function-structure studies will bring a more complete understanding of mammalian olfactory processing across spatial scales and time.

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Abstract: Friction surfacing is an emerging solid-state coating technology based on frictional heat induced plastic deformation at the tip of a consumable metallic stud that allows to deposit layers with a fine-grained recrystallized microstructure at temperatures below the melting point. The generation of sound, defect-free metallurgical joints between multiple adjacent overlapping friction surfacing deposits, also referred to as multi-track friction surfacing, from dissimilar aluminum alloys is the focus of this experimental work. An extensive volumetric defect analysis is carried out for various overlap configurations, including post-processing strategies in order to assess the inter-track bonding integrity using microscopic characterization techniques and micro-computed tomography. The effect of layer arrangement and overlap distance on the volumetric defect formation in both inter-track and layer-to-substrate interface is quantified and discussed. Post-processing via hybrid friction diffusion bonding process demonstrates a significant reduction in defect volume ratio, proving higher material efficiency. The gained knowledge was used to successfully build a multi-track multi-layer friction surfacing stack, demonstrating the suitability of this process for large-scale additive manufacturing components. The subsequent mechanical analysis reveals excellent homogeneous isotropic tensile properties of the additive structure in the range of the base material tensile strength.

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Abstract: Musculoskeletal tissues are complex hierarchical materials where mechanical response is linked to structural and material properties at different dimensional levels. Therefore, high-resolution three-dimensional tomography is very useful for assessing tissue properties at different scales. In particular, Synchrotron Radiation micro-Computed Tomography (SR-microCT) has been used in several applications to analyze the structure of bone and biomaterials. In the past decade the development of digital volume correlation (DVC) algorithms applied to SR-microCT images and its combination with in situ mechanical testing (four-dimensional imaging) have allowed researchers to visualise, for the first time, the deformation of bone tissue and its interaction with biomaterials under different loading scenarios. However, there are several experimental challenges that make these measurements difficult and at high risk of failure. Challenges relate to sample preparation, imaging parameters, loading setup, accumulated tissue damage for multiple tomographic acquisitions, reconstruction methods and data processing. Considering that access to SR-microCT facilities is usually associated with bidding processes and long waiting times, the failure of these experiments could notably slow down the advancement of this research area and reduce its impact. Many of the experimental failures can be avoided with increased experience in performing the tests and better guidelines for preparation and execution of these complex experiments; publication of negative results could help interested researchers to avoid recurring mistakes. Therefore, the goal of this article is to highlight the potential and pitfalls in the design and execution of in situ SR-microCT experiments, involving multiple scans, of musculoskeletal tissues for the assessment of their structural and/or mechanical properties. The advice and guidelines that follow should improve the success rate of this type of experiment, allowing the community to reach higher impact more efficiently.

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Abstract: Natural surface gas seeps provide a significant input of greenhouse gas emissions into the Earth’s atmosphere and hydrosphere. The gas flux is controlled by the properties of underlying fluid-escape conduits, which are present within sedimentary basins globally. These conduits permit pressure-driven fluid flow, hydraulically connecting deeper strata with the Earth’s surface; however they can only be fully resolved at sub-seismic scale. Here, a novel ‘minus cement and matrix permeability’ method using three-dimensional X-ray micro-computed tomography imaging enables the improved petrophysical linkage of outcrop and sub-surface data. The methodology is applied to the largest known outcrop of an inactive fluid-escape system, the Panoche Giant Intrusion Complex in Central California, where samples were collected along transects of the 600 to 800 m stratigraphic depth range to constrain porosity and permeability spatial heterogeneity. The presence of silica cement and clay matrix within the intergranular pores of sand intrusions are the primary control of porosity (17 to 27%) and permeability (≤1 to ca 500 mD) spatial heterogeneity within the outcrop analogue system. Following the digital removal of clay matrix and silica (opal-CT and quartz) cement derived from the mudstone host strata, the sand intrusions have porosity-permeability ranges of ca 30 to 40% and 103 to 104 mD. These calculations are closely comparable to active sub-surface systems in sedimentary basins. Field observations revealed at decreasing depth, the connected sand intrusion network reduces in thickness and becomes carbonate cemented, terminating at carbonate mounds formed from methane escape at the seafloor. A new conceptual model integrates the pore-scale calculations and field-scale observations to highlight the key processes that control sand intrusion permeability, spatially and temporally. The study demonstrates the control of matrix and cement addition on the physical properties of fluid-escape conduits, which has significance for hydrocarbon reservoir characterization and modelling, as well as subsurface CO2 and energy storage containment assessment.

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Abstract: Background: Understanding the diversity of eyes is crucial to unravel how different animals use vision to interact with their respective environments. To date, comparative studies of eye anatomy are scarce because they often involve time-consuming or inefficient methods. X-ray micro-tomography (micro-CT) is a promising high-throughput imaging technique that enables to reconstruct the 3D anatomy of eyes, but powerful tools are needed to perform fast conversions of anatomical reconstructions into functional eye models. Results: We developed a computing method named InSegtCone to automatically segment the crystalline cones in the apposition compound eyes of arthropods. Here, we describe the full auto-segmentation process, showcase its application to three different insect compound eyes and evaluate its performance. The auto-segmentation could successfully label the full individual shapes of 60-80% of the crystalline cones and is about as accurate and 250 times faster than manual labelling of the individual cones. Conclusions: We believe that InSegtCone can be an important tool for peer scientists to measure the orientation, size and dynamics of crystalline cones, leading to the accurate optical modelling of the diversity of arthropod eyes with micro-CT.