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The formation of new blood vessels and the establishment of vascular networks are crucial during brain development, in the adult healthy brain, as well as in various diseases of the central nervous system. Here, we describe a step-by-step protocol for our recently developed method that enables hierarchical imaging and computational analysis of vascular networks in postnatal and adult mouse brains. The different stages of the procedure include resin-based vascular corrosion casting, scanning electron microscopy, synchrotron radiation and desktop microcomputed tomography imaging, and computational network analysis. Combining these methods enables detailed visualization and quantification of the 3D brain vasculature. Network features such as vascular volume fraction, branch point density, vessel diameter, length, tortuosity and directionality as well as extravascular distance can be obtained at any developmental stage from the early postnatal to the adult brain. This approach can be used to provide a detailed morphological atlas of the entire mouse brain vasculature at both the postnatal and the adult stage of development. Our protocol allows the characterization of brain vascular networks separately for capillaries and noncapillaries. The entire protocol, from mouse perfusion to vessel network analysis, takes ~10 d. This protocol uses vascular corrosion casting, hierarchical synchrotron radiation microcomputed tomography imaging and computational image analysis to assess the 3D vascular network architecture in the entire postnatal and adult mouse brain.

The feasibility of dual-energy X-ray computed tomography (DECT) with experimentally implemented scatter and beam hardening corrections to quantitatively determine the 4D (3D spatial with time) evolution of partial densities of bentonite and water was examined. Compacted bentonite samples were imaged using an X-ray microtomography system with various X-ray spectra before and after two days of water infiltration. The effects of scattering and beam hardening in the projection images were corrected using beam-stop array measurements and signal-to-thickness calibration. A post-reconstruction material decomposition (MD) technique was applied to obtain the partial density distributions of bentonite and water, from which the water content distributions were subsequently derived. The results were validated by physically slicing partially saturated bentonite samples and measuring the water content of the slices gravimetrically. Additionally, a previously developed deformation measurement-based X-ray tomography method was used to derive reference results. Comparing the results from MD with those from the other techniques demonstrated that DECT can yield quantitative estimates of 4D water content distribution in bentonite with reasonable accuracy in this experimental set-up. When testing the impact of correction methods on the results, it was found that both scattering and beam hardening must be corrected, as the post-reconstruction MD is sensitive to errors in spectral measurements.

Light is a powerful and sustainable resource, but it can be detrimental to the performance and longevity of optical devices. Materials with near-zero light reflectance, i.e. superblack materials, are sought to improve the performance of several light-centered technologies. Here we report a simple top-down strategy, guided by computational methods, to develop robust superblack materials following metal-free wood delignification and carbonization (1500 °C). Subwavelength severed cells evolve under shrinkage stresses, yielding vertically aligned carbon microfiber arrays with a thickness of ~100 µm and light reflectance as low as 0.36% and independent of the incidence angle. The formation of such structures is rationalized based on delignification method, lignin content, carbonization temperature and wood density. Moreover, our measurements indicate a laser beam reflectivity lower than commercial light stoppers in current use. Overall, the wood-based superblack material is introduced as a mechanically robust surrogate for microfabricated carbon nanotube arrays. Superblack materials could innovate light-centered technologies, however, advances in superblack materials have been constrained by their complex and costly fabrication routes. Here, the authors report a simple top-down strategy, guided by computational methods, to develop robust superblack materials following metal-free wood delignification and carbonization.

In this article, we present an X-ray tomographic imaging method that is well suited for pulmonary disease studies in animal models to resolve the full pathway from gas intake to gas exchange. Current state-of-the-art synchrotron-based tomographic phase-contrast imaging methods allow for three-dimensional microscopic imaging data to be acquired non-destructively in scan times of the order of seconds with good soft tissue contrast. However, when studying multi-scale hierarchically structured objects, such as the mammalian lung, the overall sample size typically exceeds the field of view illuminated by the X-rays in a single scan and the necessity for achieving a high spatial resolution conflicts with the need to image the whole sample. Several image stitching and calibration techniques to achieve extended high-resolution fields of view have been reported, but those approaches tend to fail when imaging non-stable samples, thus precluding tomographic measurements of large biological samples, which are prone to degradation and motion during extended scan times. In this work, we demonstrate a full-volume three-dimensional reconstruction of an intact rat lung under immediate post-mortem conditions and at an isotropic voxel size of (2.75 µm)3. We present the methodology for collecting multiple local tomographies with 360° extended field of view scans followed by locally non-rigid volumetric stitching. Applied to the lung, it allows to resolve the entire pulmonary structure from the trachea down to the parenchyma in a single dataset. The complete dataset is available online (https://doi.org/10.16907/7eb141d3-11f1-47a6-9d0e-76f8832ed1b2).

In this study, cellulose microfibril (CMF) suspensions were imaged during pipe flow at consistencies of 0.4%, 1.0%, and 1.6% with optical coherence tomography (OCT) to obtain images of the structure and the local velocity of the suspension. The viscosities obtained by combining pressure loss measurement with the OCT velocity data showed typical shear thinning behavior and were in excellent agreement with viscosities obtained with ultrasound velocity profiling. The structural OCT images were used to calculate the radial and the axial floc sizes of the suspension. A fit of power law to the geometrical floc size–shear stress data gave the same power law index for all consistencies, suggesting that floc rupture dynamics is independent of consistency. The dependence of viscosity and floc size on shear stress was similar, indicating that the shear thinning behavior of CMF suspensions is closely related to the rupture dynamics of flocs. The results also showed that an apparent attenuation coefficient of the OCT signal can be used to determine the consistency of CMF suspensions.

Polarized light-based navigation in insects is facilitated by a polarization-sensitive part of the eye, the dorsal rim area (DRA). Existing methods to study the anatomy of the DRA are destructive and time-consuming. We presented a novel method for DRA localization, dissection, and measurement using 3D volumetric images from X-ray micro-computed tomography in combination with 2D photographs. Applying the method on size-polymorphic buff-tailed bumblebees, Bombus terrestris, we found that the DRA was easily obtainable from photographs of the dorsal eye region. Allometric analysis of the DRA in relation to body size in B. terrestris showed that it increased with the body size but not at the same rate. By localizing the DRA of individual bumblebees, we could also perform individual-level descriptions and inter-individual comparisons between the ommatidial structures (lens, crystalline cones, rhabdoms) of three different eye regions (DRA, non-DRA, proximate to DRA). One feature distinct to the bumblebee DRA was the smaller dimension of the crystalline cones in comparison to other regions of the eye. Using our novel methodology, we provide the first individual-level description of DRA ommatidial features and a comparison of how the DRA varies with body size in bumblebees.

Nanofibrillated cellulose (NFC) films have potential as oxygen barriers for, e.g., food packaging applications, but their use is limited by their hygroscopic characteristics. The three-dimensional microstructure of NFC films made of Pinus radiata (Radiata Pine) kraft pulp fibres has been assessed in this study, considering the structural development as a function of relative humidity (RH). The surface roughness, micro-porosity, thickness and their correlations were analyzed using X-ray microtomography (X–μCT) and computerized image analysis. The results are compared to those from scanning electron microscopy and laser profilometry. Based on a series of films having varying amounts of 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO)-mediated oxidated nanofibrils, it was demonstrated that X–μCT is suitable for assessing the surface and bulk 3D microstructure of the cellulose films. Additionally, one of the series was assessed at varying humidity levels, using the non-destructive capabilities of X–μCT and a newly developed humidity chamber for in-situ characterization. The oxygen transmission rate (OTR) of the films (20 g=m2) was below 3:7mLm-2 day-1 at humidity levels below 60% RH. However, the OTR increased considerably to 12:4mLm-2 day-1 when the humidity level increased to 80% RH. The increase in OTR was attributed to a change of the film porosity, which was reflected as an increase in local thickness. Hence, the characterization techniques applied in this study shed more light on the structures of NFC films and how they are affected by varying humidity levels. It was demonstrated that in increasing relative humidity the films swelled and the oxygen barrier properties decreased.

In solitary insect pollinators such as butterflies, sensory systems must be adapted for multiple tasks, including nectar foraging, mate-finding, and locating host-plants. As a result, the energetic investments between sensory organs can vary at the intraspecific level and even among sexes. To date, little is known about how these investments are distributed between sensory systems and how it varies among individuals of different sex. We performed a comprehensive allometric study on males and females of the butterfly Pieris napi where we measured the sizes and other parameters of sensory traits including eyes, antennae, proboscis, and wings. Our findings show that among all the sensory traits measured, only antenna and wing size have an allometric relationship with body size and that the energetic investment in different sensory systems varies between males and females. Moreover, males had absolutely larger antennae and eyes, indicating that they invest more energy in these organs than females of the same body size. Overall, the findings of this study reveal that the size of sensory traits in P. napi are not necessarily related to body size and raises questions about other factors that drive sensory trait investment in this species and in other insect pollinators in general.

Broader use of bio-based fibres in packaging becomes possible when the mechanical properties of fibre materials exceed those of conventional paperboard. Hot-pressing provides an efficient method to improve both the wet and dry strength of lignin-containing paper webs. Here we study varied pressing conditions for webs formed with thermomechanical pulp (TMP). The results are compared against similar data for a wide range of other fibre types. In addition to standard strength and structural measurements, we characterise the induced structural changes with X-ray microtomography and scanning electron microscopy. The wet strength generally increases monotonously up to a very high pressing temperature of 270 °C. The stronger bonding of wet fibres can be explained by the inter-diffusion of lignin macromolecules with an activation energy around 26 kJ mol−1 after lignin softening. The associated exponential acceleration of diffusion with temperature dominates over other factors such as process dynamics or final material density in setting wet strength. The optimum pressing temperature for dry strength is generally lower, around 200 °C, beyond which hemicellulose degradation begins. By varying the solids content prior to hot-pressing for the TMP sheets, the highest wet strength is achieved for the completely dry web, while no strong correlation was observed for the dry strength.

The formation of new blood vessels and the establishment of vascular networks are crucial during brain development, in the adult healthy brain, as well as in various diseases of the central nervous system. Here, we describe a step-by-step protocol for our recently developed method that enables hierarchical imaging and computational analysis of vascular networks in postnatal and adult mouse brains. The different stages of the procedure include resin-based vascular corrosion casting, scanning electron microscopy, synchrotron radiation and desktop microcomputed tomography imaging, and computational network analysis. Combining these methods enables detailed visualization and quantification of the 3D brain vasculature. Network features such as vascular volume fraction, branch point density, vessel diameter, length, tortuosity and directionality as well as extravascular distance can be obtained at any developmental stage from the early postnatal to the adult brain. This approach can be used to provide a detailed morphological atlas of the entire mouse brain vasculature at both the postnatal and the adult stage of development. Our protocol allows the characterization of brain vascular networks separately for capillaries and noncapillaries. The entire protocol, from mouse perfusion to vessel network analysis, takes ~10 d.