High organic carbon content constricts the potential for stable organic carbon accrual in mineral agricultural soils in Finland.

Sequestering carbon into agricultural soils is considered as a means of mitigating climate change. We used agronomic soil test results representing c. 95% of the farmed land area in Finland to estimate the potential of the uppermost 15 cm soil layer of mineral agricultural soils to sequester organic carbon (OC) and to contribute to the mitigation of climate change. The estimation of the maximum capacity of mineral matter to protect OC in stable mineral-associated form was based on the theory that clay and fine-sized (fines = clay + silt) particles have a limited capacity to protect OC. In addition, we used the clay/OC and fines/OC ratios to identify areas with a risk of erosion and reduced productivity, thus indicating priority areas potentially benefitting from the increased soil OC contents. We found that 32-40% of the mineral agricultural soils in Finland have the potential to further accumulate mineral-associated OC (MOC), while in the majority of soils, the current OC stock in the uppermost 15 cm exceeded the capacity of mineral matter to protect OC. The nationwide soil OC sequestration potential of the uppermost 15 cm in mineral agricultural soils ranged between 0.21 and 0.26 Tg, which corresponds to less than 2% of annual greenhouse gas emissions in Finland. The fields with the highest potential for SOC accrual were found in the southern and southwestern parts of the country, including some of the most intensively cultivated high-clay soils. Although the nationwide potential for additional OC sequestration was estimated to be relatively small, the current OC storage in Finnish arable mineral soils (0-15 cm) is large, 128 Tg. Farming practices enabling maximum OC input into the soil play an important role as a tool for mitigating the loss of carbon from high-OC soils in the changing climate. Furthermore, especially in high-clay areas with potential for MOC accrual, efforts to increase soil OC could help improve soil structural stability and therefore reduce erosion and the loss of nutrients to the aquatic environments.

Exposure to elevated temperature during development affects eclosion and morphology in the temperate Pieris napi butterfly (Lepidoptera: Pieridae).

Global warming has been identified as one of the main drivers of population decline in insect pollinators. One aspect of the insect life cycle that would be particularly sensitive to elevated temperatures is the developmental transition from larva to adult. Temperature-induced modifications to the development of body parts and sensory organs likely have functional consequences for adult behaviour. To date, we have little knowledge about the effect of sub-optimal temperature on the development and functional morphology of different body parts, particularly sensory organs, in ectothermic solitary pollinators such as butterflies. To address this knowledge gap, we exposed the pupae of the butterfly Pieris napi to either 23 °C or 32 °C and measured the subsequent effects on eclosion, body size and the development of the wings, proboscis, eyes and antennae. In comparison to individuals that developed at 23 °C, we found that exposure to 32 °C during the pupal stage increased mortality and decreased time to eclose. Furthermore, both female and male butterflies that developed at 32 °C were smaller and had shorter proboscides, while males had shorter antennae. In contrast, we found no significant effect of rearing temperature on wing and eye size or wing deformity. Our findings suggest that increasing global temperatures and its corresponding co-stressors, such as humidity, will impact the survival of butterflies by impairing eclosion and the proper development of body and sensory organs.

Micrometer-resolution X-ray tomographic full-volume reconstruction of an intact post-mortem juvenile rat lung.

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 ).

Laboratory phase-contrast nanotomography of unstained Bombus terrestris compound eyes.

Imaging the visual systems of bumblebees and other pollinating insects may increase understanding of their dependence on specific habitats and how they will be affected by climate change. Current high-resolution imaging methods are either limited to two dimensions (light- and electron microscopy) or have limited access (synchrotron radiation x-ray tomography). For x-ray imaging, heavy metal stains are often used to increase contrast. Here, we present micron-resolution imaging of compound eyes of buff-tailed bumblebees (Bombus terrestris) using a table-top x-ray nanotomography (nano-CT) system. By propagation-based phase-contrast imaging, the use of stains was avoided and the microanatomy could more accurately be reconstructed than in samples stained with phosphotungstic acid or osmium tetroxide. The findings in the nano-CT images of the compound eye were confirmed by comparisons with light- and transmission electron microscopy of the same sample and finally, comparisons to synchrotron radiation tomography as well as to a commercial micro-CT system were done.

NRStitcher: non-rigid stitching of terapixel-scale volumetric images.

SUMMARY: In modern microscopy, the field of view is often increased by obtaining an image mosaic, where multiple sub-images are taken side-by-side and combined post-acquisition. Mosaic imaging often leads to long imaging times that can increase the probability of sample deformation during the acquisition due to, e.g. changes in the environment, damage caused by the radiation used to probe the sample or biologically induced deterioration. Here we propose a technique, based on local phase correlation, to detect the deformations and construct an artifact-free image mosaic from deformed sub-images. The implementation of the method supports distributed computing and can be used to generate teravoxel-size mosaics. We demonstrate its capabilities by assembling a 5.6 teravoxel tomographic image mosaic of microvasculature in whole mouse brain. The method is compared to existing rigid stitching implementations designed for very large datasets, and observed to create artifact-free image mosaics in comparable runtime with the same hardware resources. AVAILABILITY AND IMPLEMENTATION: The stitching software and C++/Python source code are available at GitHub (https://github.com/arttumiettinen/pi2) along with an example dataset and user instructions.

Three-dimensional microstructural properties of nanofibrillated cellulose films.

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 3:7 mL m⁻² day⁻¹ at humidity levels below 60% RH. However, the OTR increased considerably to 12:4 mL m⁻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.

Novel Methodology for Localizing and Studying Insect Dorsal Rim Area Morphology in 2D and 3D.

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.

Wood-based superblack.

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.

Microstructural evaluation and recommendations for face masks in community use to reduce the transmission of respiratory infectious diseases.

BACKGROUND AND OBJECTIVE: Recommendations for the use of face masks to prevent and protect against the aerosols (≤5µm) and respiratory droplet particles (≥5µm), which can carry and transmit respiratory infections including severe acute respiratory syndrome coronavirus (SARS-CoV-2), have been in effect since the early stages of the coronavirus disease 2019 (COVID-19). The particle filtration efficiency (PFE) and air permeability are the most crucial factors affecting the level of pathogen transmission and breathability, i.e. wearer comfort, which should be investigated in detail. METHODS: In this context, this article presents a novel assessment framework for face masks combining X-ray microtomography and computational fluid dynamics simulations. In consideration to their widespread public use, two types of face masks were assessed: (I) two layer non-woven face masks and (II) the surgical masks (made out of a melt-blown fabric layer covered with two non-woven fabric layers). RESULTS: The results demonstrate that the surgical masks provide PFEs over 75% for particles with diameter over 0.1µm while two layer face masks are found out to have insufficient PFEs, even for the particles with diameter over 2µm (corresponding PFE is computed as 47.2%). Thus, existence of both the non-woven fabric layers for mechanical filtration and insertion of melt-blown fabric layer(s) for electrostatic filtration in the face masks were found to be highly critical to prevent the airborne pathogen transmission. CONCLUSIONS: The present framework would assist in computational assessment of commonly used face mask types based on their microstructural characteristics including fiber diameter, orientation distributions and fiber network density. Therefore, it would be also possible to provide new yet feasible design routes for face masks to ensure reliable personal protection and optimal breathability.

X-Ray Phase Contrast 3D Virtual Histology: Evaluation of Lung Alterations After Microbeam Irradiation.

PURPOSE: This study provides the first experimental application of multiscale 3-dimensional (3D) x-ray phase contrast imaging computed tomography (XPCI-CT) virtual histology for the inspection and quantitative assessment of the late-stage effects of radio-induced lesions on lungs in a small animal model. METHODS AND MATERIALS: Healthy male Fischer rats were irradiated with x-ray standard broad beams and microbeam radiation therapy, a high-dose rate (14 kGy/s), FLASH spatially fractionated x-ray therapy to avoid beamlet smearing owing to cardiosynchronous movements of the organs during the irradiation. After organ dissection, ex vivo XPCI-CT was applied to all the samples and the results were quantitatively analyzed and correlated to histologic data. RESULTS: XPCI-CT enables the 3D visualization of lung tissues with unprecedented contrast and sensitivity, allowing alveoli, vessel, and bronchi hierarchical visualization. XPCI-CT discriminates in 3D radio-induced lesions such as fibrotic scars and Ca/Fe deposits and allows full-organ accurate quantification of the fibrotic tissue within the irradiated organs. The radiation-induced fibrotic tissue content is less than 10% of the analyzed volume for all microbeam radiation therapy-treated organs and reaches 34% in the case of irradiations with 50 Gy using a broad beam. CONCLUSIONS: XPCI-CT is an effective imaging technique able to provide detailed 3D information for the assessment of lung pathology and treatment efficacy in a small animal model.

Lignin Inter-Diffusion Underlying Improved Mechanical Performance of Hot-Pressed Paper Webs.

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.

Sensory Organ Investment Varies with Body Size and Sex in the Butterfly Pieris napi.

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.

Hierarchical imaging and computational analysis of three-dimensional vascular network architecture in the entire postnatal and adult mouse brain.

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.