Optimizing illumination for full field imaging at high brilliance hard X-ray synchrotron sources.

A new technique is presented to overcome beam size limitation in full field imaging at high brilliance synchrotron sources using specially designed refractive X-ray optics. These optics defocus the incoming beam in vertical direction and reshape the intensity distribution from a Gaussian to a more desirable top-hat-shaped profile at the same time. With these optics X-ray full-field imaging of extended objects becomes possible without having to stack several scans or applying a cone beam geometry in order to image the entire specimen. For in situ experiments in general and for diffraction limited sources in particular this gain in field of view and the optimization of the intensity distribution is going to be very beneficial.

A high-spin square planar iron(ii)-siloxide and its tetrahedral allogon - structural and spectroscopic models of Fe-zeolite sites.

The famous α-Fe active sites in Fe-zeolites have recently been revealed to correspond to mononuclear high-spin iron(ii) centres in square planar coordination environments. Here we report a first iron siloxide complex which represents a faithful structural and spectroscopic model of such sites. Notably, also an allogon with a distorted structure exists and could be crystallised.

Multi-level synchrotron radiation-based microtomography of the dental alveolus and its consequences for orthodontics.

Multilevel synchrotron radiation-based microtomography has been performed on a human jaw segment obtained at autopsy by cutting increasingly smaller samples from the original segment. The focus of this study lay on the microstructure of the interface between root, periodontal ligament (PDL) and alveolar bone in order to find an answer to the question why alveolar bone remodels during orthodontic loading, when the associated stress and strain levels calculated with finite element analyses are well below the established threshold levels for bone remodeling. While the inner surface of the alveolus appears to be rather smooth on the lower resolution scans, detailed scans of the root-PDL-bone interface reveal that on a microscopical scale it is actually quite rough and uneven with bony spiculae protruding into the PDL space. Any external (orthodontic) loading applied to the root, when transferred through the PDL to the alveolar bone, will cause stress concentrations in these spiculae, rather than be distributed over a "smooth surface". As osteocyte lacunae are shown to be present in these spiculae, the local amplified stresses and strain can well be registered by the mechano-sensory network of osteocytes. In addition, a second stress amplification mechanism, due to the very presence of the lacunae themselves, is evidence that stresses and strains calculated with FE analyses, based on macroscopical scale models of teeth and their supporting structures, grossly underestimate the actual mechanical loading of alveolar bone at tissue level. It is therefore hypothesized that remodeling of alveolar bone is subject to the same biological regulatory process as remodeling in other bones.

Filling the gaps of dinosaur eggshell phylogeny: Late Jurassic Theropod clutch with embryos from Portugal.

The non-avian saurischians that have associated eggshells and embryos are represented only by the sauropodomorph Massospondylus and Coelurosauria (derived theropods), thus missing the basal theropod representatives. We report a dinosaur clutch containing several crushed eggs and embryonic material ascribed to the megalosaurid theropod Torvosaurus. It represents the first associated eggshells and embryos of megalosauroids, thus filling an important phylogenetic gap between two distantly related groups of saurischians. These fossils represent the only unequivocal basal theropod embryos found to date. The assemblage was found in early Tithonian fluvial overbank deposits of the Lourinhã Formation in West Portugal. The morphological, microstructural and chemical characterization results of the eggshell fragments indicate very mild diagenesis. Furthermore, these fossils allow unambiguous association of basal theropod osteology with a specific and unique new eggshell morphology.

Unwrapping differential x-ray phase-contrast images through phase estimation from multiple energy data.

We present a spectral phase unwrapping approach for grating-based differential phase-contrast data where the unwrapped interferometer phase shift is estimated from energy discriminated measurements using maximum likelihood principles. We demonstrate the method on tomographic data sets of a test specimen taken at different x-ray energies using synchrotron radiation. The proposed unwrapping technique was demonstrated to successfully correct the data set for phase wrapping.

Visualization of subcutaneous insulin injections by x-ray computed tomography.

We report how the three-dimensional structure of subcutaneous injections of soluble insulin can be visualized by x-ray computed tomography using an iodine based contrast agent. The injections investigated are performed ex vivo in porcine adipose tissue. Full tomography scans carried out at a laboratory x-ray source with a total acquisition time of about 1 min yield CT-images with an effective pixel size of 109 × 109 µm². The depots are segmented using a modified Chan-Vese algorithm and we are able to observe differences in the shape of the injection depot and the position of the depot in the skin among equally performed injections. To overcome the beam hardening artefacts, which affect the quantitative prediction of the volume injected, we additionally present results concerning the visualization of two injections using synchrotron radiation. The spatial concentration distribution of iodine is calculated to show the dilution of the insulin drug inside the depot. Characterisation of the shape of the depot and the spatial concentration profile of the injected fluid is important knowledge when improving the clinical formulation of an insulin drug, the performance of injection devices and when predicting the effect of the drug through biomedical simulations.

In vivo corrosion and corrosion protection of magnesium alloy LAE442.

The aim of this study was to investigate whether the extruded magnesium alloy LAE442 reacts in vivo with an appropriate host response and to investigate how an additional magnesium fluoride (MgF(2)) coating influences the in vivo corrosion rate. Forty cylinders were machined from extruded LAE442 and 20 of these were coated additionally with MgF(2) and implanted into the medial femur condyle of adult rabbits. Synchrotron-radiation-based X-ray computed micro-tomography (SRmicroCT) was used to quantitatively analyse corrosion non-destructively in vivo and comparisons were made to magnesium degradation rates based on area measurements of the remaining metal on uncalcified sections. Blood concentrations of the alloying elements were measured below toxicological limits. The MgF(2) layer was no longer detected after 4 weeks of implantation by particle-induced gamma emission, and the MgF(2) coating reduced the blood content of alloying elements during the first 6 weeks of implantation with no elevated fluoride concentration in the adjacent bone. Histopathological examinations of liver showed in 9 out of 40 cases minimal infiltrations of heterophil granulocytes of unknown origin (5 LAE442, 4 LAE442+MgF(2)). The kidneys were mainly regular in structure. The synovial tissue showed a granular cell infiltration as a temporary observation in the LAE442+MgF(2) group after 2 weeks. No subcutaneous gas cavities were observed clinically and on postoperative X-rays in all animals. All specimens were scanned by SRmicroCT at 2, 4, 6 and 12 weeks postoperatively before uncalcified sections were performed. All magnesium implants have been observed in direct bone contact and without a fibrous capsule. Localized pitting corrosion occurred in coated and uncoated magnesium implants. This study shows that the extruded magnesium alloy LAE442 provides low corrosion rates and reacts in vivo with an acceptable host response. The in vivo corrosion rate can be further reduced by additional MgF(2) coating.

High-resolution X-ray tomography of the human inner ear: synchrotron radiation-based study of nerve fibre bundles, membranes and ganglion cells.

The combination of osmium tetroxide staining and high-resolution tomographic imaging using monochromatic X rays allows visualizing cellular structures of the human inner ear, that is, the organ of Corti, the stria vascularis and further soft tissues of the membranous labyrinth, in three-dimensional space with isotropic micrometre resolution. This approach permits to follow the course of nerve fibre bundles in a major part of the specimen and reveals the detailed three-dimensional arrangement of individual ganglion cells with distinct nuclei by means of X-ray tomography for the first time. The non-destructive neuron cell counting in a selected volume of 125 microm x 800 microm x 600 microm = 0.06 mm(3) gives rise to the estimate that 2000 ganglion cells are present along 1 mm organ of Corti.

Synchrotron radiation-based microtomography of alveolar support tissues.

OBJECTIVES: To study the alveolar support structures using synchrotron radiation (SR)-based microtomography with particular focus on the alveolar surface. DESIGN: High-resolution microtomography of jaw segments of various species and subsequent three-dimensional (3D) reconstruction. SETTING AND SAMPLE POPULATION: Microtomography was performed at the DORIS-ring of the synchrotron facility of HASYLAB/DESY in Hamburg, Germany. The samples consisted of human, simian and porcine jaw segments. RESULTS: With SR being monochromatic, no beam-hardening artifacts could occur and the grey values in the scans were therefore directly related to the local tissue densities. Apart from the mineralized tissues, the beam energy was low enough to allow for the visualization of soft tissues like the fibers of the periodontal ligament (PDL) and blood vessels. 3D reconstructions of the alveolar bone showed that it can be rough and sharply edged. Furthermore, an intricate network of marrow cavities and blood vessels penetrates its surface. Differences in the local grey value distribution in the alveolar bone pointed to remodeling activity in the close vicinity of the PDL. CONCLUSION: The assumption that the alveolar bone surface is smooth and continuous is not correct. This means that even small orthodontic loads can already give rise to high local stresses and strains in the bone and thus initiate remodeling processes.

Inhomogeneous plastic flow investigated by X-ray absorption microtomography of an aluminium alloy containing marker particles.

By combining a synchrotron X-ray source and the microtomography technique, the displacement gradient tensor components can be measured directly throughout the volume of a metal sample containing highly absorbing marker particles to detect material flow. The present article describes application of these techniques to compression of a rectangular Al-W specimen with a central hole. The results show that the externally imposed axial displacement gradient is accommodated mainly in two 45 degrees shear bands passing through the hole. A simple deformation analysis provides insight into shear band formation.

Comparison of microfocus- and synchrotron X-ray tomography for the analysis of osteointegration around Ti6Al4V implants.

Micro-computed tomography (microCT) provides quantitative three-dimensional information of bone around titanium implants similar to classical histology. The study, based on an animal model, containing cuboid-shaped biofunctionalised Ti6Al4V implants with surrounding bone after 4 weeks, is performed using 3 microCT-systems with X-ray tubes, one synchrotron-radiation-based microCT-system (SRmicroCT), and classical histology. Although the spatial resolution of the microCT-systems is comparable, only the results of SRmicroCT agree with results of classical histology. The X-ray tube sources give rise to huge artefacts in the tomograms (interface scattering, beam hardening), which impaired the quantitative analysis of bone up to about 200microm from the implant surface. Due to the non-destructive character of microCT the specimens can be subsequently examined by classical histology without restriction. The quantitative comparison of bone formation uncovers the strong dependence of the newly formed bone from the selected slice. This implies the necessity of 3D analysis. SRmicroCT and classical histology prove that surface modifications of the titanium implant significantly influence the bone formation. Using SRmicroCT, the preparation artefacts due to cutting and polishing are excluded.

A novel method to produce hydroxyapatite objects with interconnecting porosity that avoids sintering.

Porous objects of carbonated apatite were prepared by mixing polyvinyl alcohol fibres (PVA) and sodium chloride as porogens with nanocrystalline carbonated apatite powder. After cold isostatic pressing (CIP) and dissolving the porogens, the bioceramic showed an interconnecting porosity with pore diameters in the range of 250-400 microm. The method can be transposed to any material that is insoluble in water (like many polymers). Such objects are promising for bone regeneration because the interconnecting porosity in carbonated apatite provides a good environment for bone attachment and ingrowth.

Multiple-beam X-ray interferometry for phase-contrast microtomography.

The first successful operation of an X-ray interferometer under conditions of non-planar three-beam diffraction for phase-contrast X-ray microtomography is reported. Intrinsic phase differences of the reflections used cancel from the three-dimensional phase image of the specimen. With simultaneous hkl and hkl reflections of a synchrotron radiation beam in a side-by-side geometry, the size of the usable field of view is doubled and the investigated specimen volume is increased by a factor of four. As an example, the reconstructed slice of a mouse kidney is shown in phase contrast at 71 keV. Optimized choices of three-beam reflections and matching interferometer geometries useful for applications are presented.

Three-dimensional imaging of nerve tissue by x-ray phase-contrast microtomography.

We show that promising information about the three-dimensional (3D) structure of a peripheral nerve can be obtained by x-ray phase-contrast microtomography (p-microCT; Beckmann, F., U. Bonse, F. Busch, and O. Günnewig, 1997. J. Comp. Assist. Tomogr. 21:539-553). P-microCT measures electronic charge density, which for most substances is proportional to mass density in fairly good approximation. The true point-by-point variation of density is thus determined in 3D at presently 1 mg/cm3 standard error (SE). The intracranial part of the rat trigeminal nerve analyzed for the presence of early schwannoma "microtumors" displayed a detailed density structure on p-microCT density maps. The average density of brain and nerve tissue was measured to range from 0.990 to 0.994 g/cm3 and from 1.020 to 1.035 g/cm3, respectively. The brain-nerve interface was well delineated. Within the nerve tissue, a pattern of nerve fibers could be seen that followed the nerve axis and contrasted against the bulk by 7 to 10 mg/cm3 density modulation. Based on the fact that regions of tumor growth have an increased number density of cell nuclei, and hence of the higher z element phosphorus, it may become possible to detect very early neural "microtumors" through increases of average density on the order of 10 to 15 mg/cm3 by using this method.

X-ray microtomography (microCT) using phase contrast for the investigation of organic matter.

PURPOSE: We show that microtomography (microCT) using synchrotron radiation (SR) can be extended to include X-ray phase contrast, which is two to three orders of magnitude more sensitive than conventional attenuation contrast and better suited for the investigation of specimens consisting chiefly of light elements for photon energies ranging at least from 1 to 100 keV. METHOD: Phase contrast is generated by placing the specimen in one of the interfering beams of an X-ray interferometer. With use of 12-keV X-rays, phase projections of the specimen are recorded at 180 or 360 angular settings equally spaced between 0 and 180 degrees. One phase projection consists of four pairs of "associated" radiograms in the sense that one is taken with and the other without the specimen in the beam. Between pairs a parallel-sided phase-shifter plate is rotated for changing the relative phase of the two interfering beams by multiples of pi/2 rad. By calculating phase-weighted sums of all associated pairs of radiograms, true phase-shift projections are obtained for all angular settings of the specimen, which are then reconstructed. RESULTS: Three-dimensional images have been obtained from rat cerebrum and rat trigeminal nerve, showing cell structures at 8- to 15-micron spatial resolution. Gray and white matter of cerebrum and neurons in the trigeminal nerve are clearly visible. CONCLUSION: X-ray phase-contrast microCT is becoming a valuable tool for studies of organic samples in medicine and biology.

[New possibilities for structural analysis of bone biopsies using microcomputer tomography (muCT)]. / Neue Möglichkeiten der Strukturanalyse von Knochenbiopsien bei Anwendung der Mikrocomputertomographie (muCT).

The combined histological and microcomputed analysis of human iliac crest biopsies leads to major advances in our understanding of three-dimensional bone architecture. Microcomputed tomography avoids the time-consuming reconstruction and artifacts of serial sections. Furthermore, its high resolution allows the recording of structural differences as low as 10 microns. Thus, three-dimensional analysis in combination with histological evaluation of cellular dynamics facilitates earlier and easier recording of changes of cancellous bone.

3D computed X-ray tomography of human cancellous bone at 8 microns spatial and 10(-4) energy resolution.

Human cancellous bone was imaged and its absorptive density accurately measured in three dimensions (3D), nondestructively and at high spatial resolution by means of computerized microtomography (microCT). Essential for achieving the resolution and accuracy was the use of monoenergetic synchrotron radiation (SR) which avoided beam hardening effects, secured excellent contrast conditions including the option of energy-modulated contrast, and yet provided high intensity. To verify the resolution, we selected objects of approximately 8 micron size that could be observed on tomograms and correlated them in a unique manner to their counter images seen in histological sections prepared from the same specimen volume. Thus we have shown that the resolution expected from the voxel size of 8 microns used in the microCT process is in effect also attained in our results. In achieving the present results no X-ray-optical magnification was used. From microCT studies of composites (Bonse et al., X-ray tomographic microscopy (XTM) applied to carbon-fibre composites. In: Materlik G, ed. HASYLAB Jahresbericht 1990. Hamburg: DESY, 1990; 567-568) we know that by including X-ray magnification a resolution below 2 microns is obtained. Therefore, with foreseeable development of our microCT method, the 3D and nondestructive investigation of structures in mineralized bone on the 2 micron level is feasible. For example, it should be possible to study tomographically the 3D distribution and amount of osteoclastic resorption in the surrounding bone structure.