Phase-contrast X-ray Microtomography Research Articles

Characterization of spring and durum wheat using non-destructive synchrotron phase contrast X-ray microtomography during storage

Post-harvest losses during cereal grain storage are a big concern in both developing and developed countries, where spring and durum wheat are staple food grains. Varieties under these classes behave differently under storage, which affects their end storage life. High resolution imaging data of dry as well as spoiled seed are not available for any class of wheat; therefore, an attempt was made to generate 3D data for better understanding of seed structure and changes due to spoilage. Six wheat varieties (3 varieties for each class of wheat) were stored for 5 week at 17% moisture content (wb) before scanning. Seeds were also stored in a freezer (-18 °C) for further scanning to determine if any changes occur in the structure of seeds due to freezing. Spring varieties of wheat performed better than durum varieties and freezing did not affect seed structure. Data could also help plant breeders to develop varieties that do not easily spoil, adjust grain processing techniques, and develop post-harvest recommendations for other wheat varieties.

Micro- and nanostructure specific X-ray tomography reveals less matrix formation and altered collagen organization following reduced loading during Achilles tendon healing

Recovery of the collagen structure following Achilles tendon rupture is poor, resulting in a high risk for re-ruptures. The loading environment during healing affects the mechanical properties of the tendon, but the relation between loading regime and healing outcome remains unclear. This is partially due to our limited understanding regarding the effects of loading on the micro- and nanostructure of the healing tissue. We addressed this through a combination of synchrotron phase-contrast X-ray microtomography and small-angle X-ray scattering tensor tomography (SASTT) to visualize the 3D organization of microscale fibers and nanoscale fibrils, respectively. The effect of in vivo loading on these structures was characterized in early healing of rat Achilles tendons by comparing full activity with immobilization. Unloading resulted in structural changes that can explain the reported impaired mechanical performance. In particular, unloading led to slower tissue regeneration and maturation, with less and more disorganized collagen, as well as an increased presence of adipose tissue. This study provides the first application of SASTT on soft musculoskeletal tissues and clearly demonstrates its potential to investigate a variety of other collagenous tissues. Statement of significanceCurrently our understanding of the mechanobiological effects on the recovery of the structural hierarchical organization of injured Achilles tendons is limited. We provide insight into how loading affects the healing process by using a cutting-edge approach to for the first time characterize the 3D micro- and nanostructure of the regenerating collagen. We uncovered that, during early healing, unloading results in a delayed and more disorganized regeneration of both fibers (microscale) and fibrils (nanoscale), as well as increased presence of adipose tissue. The results set the ground for the development of further specialized protocols for tendon recovery.

Distribution of corpora amylacea in the human midbrain: using synchrotron radiation phase-contrast microtomography, high-field magnetic resonance imaging, and histology.

Corpora amylacea (CA) are polyglucosan aggregated granules that accumulate in the human body throughout aging. In the cerebrum, CA have been found in proximity to ventricular walls, pial surfaces, and blood vessels. However, studies showing their three-dimensional spatial distribution are sparse. In this study, volumetric images of four human brain stems were obtained with MRI and phase-contrast X-ray microtomography, followed up by Periodic acid Schiff stain for validation. CA appeared as hyperintense spheroid structures with diameters up to 30 μm. An automatic pipeline was developed to segment the CA, and the spatial distribution of over 200,000 individual corpora amylacea could be investigated. A threefold-or higher-density of CA was detected in the dorsomedial column of the periaqueductal gray (860-4,200 CA count/mm3) than in the superior colliculus (150-340 CA count/mm3). We estimated that about 2% of the CA were located in the immediate vicinity of the vessels or in the peri-vascular space. While CA in the ependymal lining of the cerebral aqueduct was rare, the sub-pial tissue of the anterior and posterior midbrain contained several CA. In the sample with the highest CA density, quantitative maps obtained with MRI revealed high R2* values and a diamagnetic shift in a region which spatially coincided with the CA dense region.

Exploiting synchrotron X-ray tomography for a novel insight into flax-fibre defects ultrastructure

Flax fibres are valuable reinforcements for tomorrow’s composites. However, defects called kink-bands, which mainly appear on fibres during the extraction and transformation phases, might affect their mechanical properties. Defects induced pores within the kink-band are investigated in this work. They were morphologically explored using synchrotron phase-contrast X-ray microtomography, a technique that displays a sharp 3D representation of the pores. The study highlights the link between kink-bands and secondary cell wall ultrastructure. Pores are organised concentrically around the lumen, and their low thickness suggest that they are located at the interface between cellulose layers within S2 (G) layer. Moreover, the pores inclination with reference to the lumen axis follows the typical microfibrillar angle changes observed in the literature in the kink-band region. The volumes of the pores were measured, and a local increase in porosity was revealed in zones where defects are most severe along the fibre.

New Growth-Related Features of Wheat Grain Pericarp Revealed by Synchrotron-Based X-ray Micro-Tomography and 3D Reconstruction.

Wheat (Triticum aestivum L.) is one of the most important crops as it provides 20% of calories and proteins to the human population. To overcome the increasing demand in wheat grain production, there is a need for a higher grain yield, and this can be achieved in particular through an increase in the grain weight. Moreover, grain shape is an important trait regarding the milling performance. Both the final grain weight and shape would benefit from a comprehensive knowledge of the morphological and anatomical determinism of wheat grain growth. Synchrotron-based phase-contrast X-ray microtomography (X-ray µCT) was used to study the 3D anatomy of the growing wheat grain during the first developmental stages. Coupled with 3D reconstruction, this method revealed changes in the grain shape and new cellular features. The study focused on a particular tissue, the pericarp, which has been hypothesized to be involved in the control of grain development. We showed considerable spatio-temporal diversity in cell shape and orientations, and in tissue porosity associated with stomata detection. These results highlight the growth-related features rarely studied in cereal grains, which may contribute significantly to the final grain weight and shape.

Micro- and nano-tomography analysis of mouse soleus muscle using radiation.

In this study, we analyze radiation images of muscle structure of mice soleus muscles using radiation source-based microtomography and nanotomography. Soleus muscle samples were collected for analysis from 8-week-old male Institute of Cancer Research mice. First, phase-contrast X-ray microtomography was employed in these experiments. Then to obtain images with excellent contrast, imaging was performed using monochromatic light with excellent transmission power. To analyze additional muscle structures in higher magnification images than these images, nanotomography was performed, which facilitated obtaining high-magnification and high-resolution images. Muscle tissue microstructures were confirmed through three-dimensional images obtained from phase-contrast X-ray microtomography. Thus, the muscle tissue's overall shape at microscopic level can be captured. Additionally, a single muscle fiber was examined using hard X-ray nano-imaging, through which we could observe the alignment of countless myofibrils, that is, actin and myosin filaments in the muscle fibers. Thus, the methodology adopted here proved to be advantageous in analyzing the muscle tissue's overall structure with microtomography and in observing the myofibrils in detail using nanotomography.

Efficient three-dimensional characterization of C/C composite reinforced with densely distributed fibers via X-ray phase-contrast microtomography

Carbon fiber (CF)/pyrolytic graphite (PG) composites are promising structural materials for molten salt reactors because of their superior performance. Due to the minor density difference between CF and PG, existing methods are impractical for efficient three-dimensional characterization of CF/PG composites. Therefore, in this study, a method based on in-line phase-contrast X-ray microtomography was developed to solve the aforementioned problem. Experimental results demonstrate that the method is suitable for comprehensive characterization of CF/PG composites. The relationship between the microporous defects and fiber orientations of such composites was also elucidated. The findings can be useful for improving the manufacturing process of CF/PG composites.

Comprehensive characterization of TSV etching performance with phase-contrast X-ray microtomography

Comprehensive evaluation of through-silicon via (TSV) reliability often requires deterministic and 3D descriptions of local morphological and statistical features of via formation with the Bosch process. Here, a highly sensitive phase-contrast X-ray microtomography approach is presented based on recorrection of abnormal projections, which provides comprehensive and quantitative characterization of TSV etching performance. The key idea is to replace the abnormal projections at specific angles in principles of linear interpolation of neighboring projections, and to distinguish the interface between silicon and air by using phase-retrieval algorithms. It is demonstrated that such a scheme achieves high accuracy in obtaining the etch profile based on the 3D microstructure of the vias, including diameter, bottom curvature radius, depth and sidewall angle. More importantly, the 3D profile error of the via sidewall and the consistency of parameters among all the vias are achieved and analyzed statistically. The datasets in the results and the 3D microstructure can be applied directly to a reference and model for further finite element analysis. This method is general and has potentially broad applications in 3D integrated circuits.

The oldest fossil darkling beetle of the genus Neomida Latreille, 1829 (Coleoptera: Tenebrionidae) from Eocene Baltic amber examined with X-ray microtomography.

Based on a single well-preserved specimen from Eocene Baltic amber, a new tenebrionid beetle Neomida groehni Nabozhenko et Bukejs sp. nov. (Diaperinae: Diaperini) is described and illustrated using phase-contrast X-ray microtomography. This oldest representative of the genus differs from all known extant and extinct congeners by the dorsally very weakly convex pronotum with undulate lateral margins (while other Neomida have a pronotum that is strongly convex along its transverse axis, with evenly rounded lateral margins); distinct, right posterior angles of pronotum; and elevated sutural area of the elytra.

Cretaciella sorianoae gen. et sp. nov. (Coleoptera, Leiodidae, Cholevinae, Oritocatopini), Anophthalmic Species from Albian Amber of the Escucha Formation (Alava, Spain)

Cretaciella sorianoae gen. et sp. nov. (Coleoptera, Leiodidae, Cholevinae, Oritocatopini) from Albian amber of the Escucha Formation (Alava, Spain) is described and illustrated. This is the first species of Leiodidae from Alava amber and the first Cholevinae from Cretaceous amber. External and internal structures are investigated by propagation phase contrast X-ray microtomography. Based on both external and genital structures, Cretaciella is tentatively placed in the tribe Oritocatopini, the extant species of which occur in Sub-Saharan Africa. This specimen has no visible eyes or hind wings, which suggests an adaptation to subterranean or at least to soil litter environments. The biogeography of the tribe is succinctly discussed.

Study of drug particle distributions within mini-tablets using synchrotron X-ray microtomography and superpixel image clustering.

Uniform drug distribution within fast disintegrating tablets is a key quality measure to ensure a reliable, steady, and targeted release of the contained active pharmaceutical ingredients. In this work, the drug particle distribution in mini-tablets was studied with synchrotron phase contrast X-ray microtomography. Mini-tablets had a weight of 9.5mg and a drug load from 2.5% to 20%. Moxidectin, a drug used for treatment of parasitic infections, was used as a model compound. Drug content covered a range from 91% to 121% of the target dose. A linear iterative clustering (SLIC) superpixel method was used for segmentation, analysis, and visualization of the spatial distribution of individual tablet components (i.e., pores, excipients, and drug). Results show that the drug was not uniformly distributed within the tablet, revealing an increasing drug load towards the tablets' outer boundaries and thus indicative of a radial displacement of drug particles during compaction. The presented method can be used for the quantitative analysis of drug content and drug distribution within pharmaceutical tablets, allowing for the optimization of fast disintegrating formulations. The results also affirm that that drug loads up to 20% will not lead to segregation for moxidectin.

Ultrastructural Analysis of Human Gallstones using Synchrotron Radiation µCT.

Gallstone formation is a pathological process of mineralization in the human body. Determination of the morphology and ultrastructure of gallstones holds the key to understanding the pathophysiology of gallbladder disease. Synchrotron radiation phase-contrast Xray microtomography is a novel technology, which is designed for comprehensive analysis of gallstone ultrastructure. Nine human gallstones were obtained from the Department of Pathology, Qingpu branch of Zhongshan Hospital Affiliated to Fudan University (China), and scanned by synchrotron radiation µCT (SR µCT). The imaging data generated by SR µCT scan were analyzed. The three-dimensional ultrastructure of human gallstones corresponding to their cholesterol and bile pigment composition was determined. The ultrastructure of gallstones exhibits considerable diversity and complexity. The synchrotron radiation phase-contrast X-ray microtomography is a valuable tool for in-depth study of human gallstones.

Lilioceris groehni sp. n.: the first authentic species of Criocerinae (Coleoptera, Chrysomelidae) from Baltic amber

Based on a single well-preserved specimen from Eocene Baltic amber, Lilioceris groehni sp. n. is described and illustrated using phase-contrast X-ray microtomography. It is the first described species of Criocerinae (Coleoptera: Chrysomelidae) from Baltic amber. A check-list of fossil Criocerinae is provided. Placement of Crioceris pristiana (Germar, 1813) is discussed, this species is removed from Criocerinae and placed in Coleoptera incertae sedis.

Assessment of the 3 D Pore Structure and Individual Components of Preshaped Catalyst Bodies by X-Ray Imaging.

Porosity in catalyst particles is essential because it enables reactants to reach the active sites and it enables products to leave the catalyst. The engineering of composite-particle catalysts through the tuning of pore-size distribution and connectivity is hampered by the inability to visualize structure and porosity at critical-length scales. Herein, it is shown that the combination of phase-contrast X-ray microtomography and high-resolution ptychographic X-ray tomography allows the visualization and characterization of the interparticle pores at micro- and nanometer-length scales. Furthermore, individual components in preshaped catalyst bodies used in fluid catalytic cracking, one of the most used catalysts, could be visualized and identified. The distribution of pore sizes, as well as enclosed pores, which cannot be probed by traditional methods, such as nitrogen physisorption and isotherm analysis, were determined.

Propagation-based phase-contrast X-ray microtomography of a cerebral protection device retrieved after carotid artery stenting

Phase-contrast synchrotron X-ray microtomography (pcSyncX) based on the highly coherent X-ray beam has previously been used to visualize the microstructures of biologic specimens, but it has never been used to evaluate embolic debris adherent on a cerebral protection device (CPD). The purpose of this study was to demonstrate the feasibility of pcSyncX for evaluating embolic debris during carotid artery stenting (CAS). Five patients (four males, age range 67-77 years) with severe carotid artery stenosis underwent CAS. The retrieved CPD was exposed to synchrotron radiation and 1000 pcSyncX projection images were obtained by rotating the CPD through 180°. An X-ray shadow of a CPD was converted into a visual image by the scintillator. After microtomographic reconstruction, the three-dimensionally reconstructed images were further segmented into the embolic debris and CPD. The total volume of emboli was calculated by summing the volume at each scanning level. The number of membrane pores covered by emboli as seen from the outer surface was counted and the percentage of covered area was calculated. Embolic debris was clearly demonstrated not only on the inner surface and within pores but also on the outer surface of the CPD. The mean total volume of embolic debris was 0.538 × 10(-6) mm(3) (range 0.225-0.965 × 10(-6) mm(3)). Most (61.5%) of the debris was located at the apical one-third of the CPD and 20.8% of the pore area was covered by debris.

Three-dimensional phase-contrast X-ray microtomography with scanning–imaging X-ray microscope optics

A three-dimensional (3D) X-ray tomographic micro-imaging system has been developed. The optical system is based on a scanning-imaging X-ray microscope (SIXM) optics, which is a hybrid system consisting of a scanning microscope optics with a one-dimensional (1D) focusing (line-focusing) device and an imaging microscope optics with a 1D objective. In the SIXM system, each 1D dataset of a two-dimensional (2D) image is recorded independently. An object is illuminated with a line-focused beam. Positional information of the region illuminated by the line-focused beam is recorded with the 1D imaging microscope optics as line-profile data. By scanning the object with the line focus, 2D image data are obtained. In the same manner as for a scanning microscope optics with a multi-pixel detector, imaging modes such as phase contrast and absorption contrast can be arbitrarily configured after the image data acquisition. By combining a tomographic scan method and the SIXM system, quantitative 3D imaging is performed. Results of a feasibility study of the SIXM for 3D imaging are shown.

Simple Scanning Phase-Contrast X-ray Tomography Using Intensity Detectors

We have developed a phase-contrast X-ray microtomography system based on scanning microscopy with X-ray focusing optics. The system is very simple because only two conventional intensity detectors and a half-obstructing-plate for the second detector are installed into the conventional scanning microscopy setup. We constructed the system with a 450-nm-size focused beam of a 10 keV X-ray. The system is operated with a sampling rate of 100 Hz. Adopting data correction using a guide object and data interpolation procedure in sinograms, tomographic reconstruction images with a high spatial resolution and phase sensitivity are obtained.

Phase contrast X-ray microtomography of the Rhodnius prolixus head: Comparison of direct reconstruction and phase retrieval approach

We have used phase-contrast X-ray microtomography (PPC-μCT) to study the head of the blood-feeding bug, Rhodnius prolixus, which is one of the most important insect vector of Trypanosoma cruzi, ethiologic agent of Chagas disease in Latin America. Images reconstructed from phase-retrieved projections processed by ANKA phase are compared to those obtained through direct tomographic reconstruction of the flat-field-corrected transmission radiographs. It should be noted that the relative locations of the important morphological internal structures are observable with a precision that is difficult to obtain without the phase retrieval approach.

First fossil record of polypore fungus beetles from Lower Cretaceous amber of France

The first fossil genus and species of Tetratomidae (Coleoptera) is described, from the Lower Cretaceous amber deposits of France. The new genus represents the first insect to be recognized inside an opaque piece of amber, through the use of propagation phase contrast X-ray microtomography using synchrotron radiation. This new finding proves the capabilities of this imaging technique in amber inclusions, as well as increases the knowledge of fossil tenebrionoids, a group scarcely recognized in the Cretaceous fossil record.

Phase-contrast X-ray microtomography of mouse fetus

SummaryA phase-contrast X-ray microtomography system using the Talbot imaging has been built at the SPring-8 synchrotron radiation facility. This system has much higher density resolution than absorption-based X-ray microtomography. The tomographic sections of formalin-fixed mouse fetuses obtained with this method clearly depict various organs without any staining at a pixel resolution of up to 5 µm. Since this technique allows us to obtain three-dimensional structural information without sectioning, it will be particularly useful to examine anomalies that take place during development. It can be also used to quantitatively measure volume and mass of organs during development.