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X-ray phase-contrast techniques are powerful methods for discerning features with similar densities, which are normally indistinguishable with conventional absorption contrast. While these techniques are well-established tools at large-scale synchrotron facilities, efforts have increasingly focused on implementations at laboratory sources for widespread use. X-ray speckle-based imaging is one of the phase-contrast techniques with high potential for translation to conventional x-ray systems. It yields phase-contrast, transmission, and dark-field images with high sensitivity using a relatively simple and cost-effective setup tolerant to divergent and polychromatic beams. Recently, we have introduced the unified modulated pattern analysis (UMPA) [Phys. Rev. Lett. 118, 203903 (2017)PRLTAO0031-900710.1103/PhysRevLett.118.203903], which further simplifies the translation of x-ray speckle-based imaging to low-brilliance sources. Here, we present the proof-of-principle implementation of UMPA speckle-based imaging at a microfocus liquid-metal-jet x-ray laboratory source.

X-ray grating interferometry (XGI) is a phase-contrast imaging technique that allows for a quantitative measurement of the refractive index with high density resolution in a model-independent manner—i.e. without a priori knowledge of the specimen composition. However, the retrieval of the X-ray wavefront phase shift relies on the accurate measurement of the interference pattern phase shift, making XGI vulnerable to phase wrapping when the interference pattern phase shift, related to the derivative of the wavefront phase shift, is large. Standard procedure for avoiding phase wrapping involves submerging the specimen in a water bath to reduce the mismatch of the index of refraction at the boundaries, but this requires a top-down rotation stage and is susceptible to gas bubble formation inside the water bath. Our team has presented an algorithm to remove phase wrapping artifacts for cylindrically shaped specimens that is applied to the phase-retrieved sinogram. This algorithm models and replaces phase-wrapped data to prevent the spread of “cupping” artifacts due to the integration of the differential phase during reconstruction. We give a criterion for selecting the modeling parameters so that the resulting measurement of the index of refraction matches the results of measurements without phase wrapping. We also apply this technique to cases where phase wrapping occurs at multiple interfaces. This algorithm allows for XGI measurements without a water bath and top-down rotation stage at synchrotron and laboratory facilities, especially as sensitivity increases.

Phase-contrast micro-tomography using synchrotron radiation has yielded superior soft tissue visualization down to the sub-cellular level. The isotropic spatial resolution down to about one micron is comparable to the one of histology. The methods, however, provide different physical quantities and are thus complementary, also allowing for the extension of histology into the third dimension. To prepare for cross-sectional animal studies on epilepsy, we have standardized the specimen’s preparation and scanning procedure for mouse brains, so that subsequent histology remains entirely unaffected and scanning of all samples (n = 28) is possible in a realistic time frame. For that, we have scanned five healthy and epileptic mouse brains at the ID19 beamline, ESRF, Grenoble, France, using grating- and propagation-based phase contrast micro-tomography. The resulting datasets clearly show the cortex, ventricular system, thalamus, hypothalamus, and hippocampus. Our focus is on the latter, having planned kainate-induced epilepsy experiments. The cell density and organization in the dentate gyrus and Ammon’s horn region were clearly visualized in control animals. This proof of principle was required to initiate experiment. The resulting three-dimensional data have been correlated to histology. The goal is a brain-wide quantification of cell death or structural reorganization associated with epilepsy as opposed to histology alone that represents small volumes of the total brain only. Thus, the proposed technique bears the potential to correlate the gold standard in analysis with independently obtained data sets. Such an achievement also fuels interest for other groups in neuroscience research to closely collaborate with experts in phase micro-tomography.

Computed laminography (CL), a long established NDT method, is ideal for the non-destructive evaluation of plate-like structures, for which conventional computed tomography (CT) is less well suited. This paper demonstrates the feasibility of implementing raster CL on a Nikon Metrology custom build X-ray CT scanner without the need for specialised equipment. To provide a more principled way to demonstrate the widespread adoption of CL for defect characterization in plate-like structures, a test specimen was designed and manufactured from Polyoxymethylene (Delrin) that allows well defined features of different size and orientation to be characterized. The study highlights the relation between direction-dependent spatial resolution, feature geometry and scan trajectory. It demonstrates that high-resolution and high-speed in situ volume-imaging of plate-like structures with large aspect ratios is viable at levels that would be challenging via CT or alternative three-dimensional NDT methods.

Hard X-ray tomography with Paganin's widespread single-distance phase retrieval filter improves contrast-to-noise ratio (CNR) while reducing spatial resolution (SR). We demonstrate that a Gaussian filter provided larger CNR at high SR with interpretable density measurements for two medically relevant soft tissue samples. Paganin's filter produced larger CNR at low SR, though \emph{a priori} assumptions were generally false and image quality gains diminish for CNR $>1$. Therefore, simple absorption measurements of low-$Z$ specimens combined with Gaussian filtering can provide improved image quality and model-independent density measurements compared to single-distance phase retrieval., This article may be downloaded for personal use only. Any other use requires prior permission of the author and AIP Publishing. This article appeared in Applied Physics Letters 116, 023702 (2020), and may be found at https://aip.scitation.org/doi/10.1063/1.5133742

X-ray phase-contrast and dark-field imaging provides valuable, complementary information about the specimen under study. Among the multimodal X-ray imaging methods, X-ray grating interferometry and speckle-based imaging have drawn particular attention, which, however, in their common implementations incur certain limitations that can restrict their range of applications. Recently, the unified modulated pattern analysis (UMPA) approach was proposed to overcome these limitations and combine grating- and speckle-based imaging in a single approach. Here, we demonstrate the multimodal imaging capabilities of UMPA and highlight its tunable character regarding spatial resolution, signal sensitivity and scan time by using different reconstruction parameters., Journal of Instrumentation, 13, ISSN:1748-0221

Our body is hierarchically organized down to individual cells. Cutting-edge clinical imaging facilities reach a spatial resolution of a fraction of a millimeter, living cells invisible. A decade ago, post-mortem X-ray imaging by means of synchrotron radiation enabled the identification of Os-stained ganglion and unstained Purkinje cells. Very recently, even sub-cellular structures, such as nucleolus and the dendritic tree of Purkinje cells, were extracted by means of phase-contrast single-distance synchrotron radiation-based hard X-ray tomography. At the same time, conventional absorption-contrast, laboratory-based micro computed tomography was successfully applied to visualize brain components including individual Purkinje cells within a cerebellum specimen. Thus, the goal of isotropic-cellular-resolution visualization of soft tissues within a laboratory environment without application of any dedicated contrast agent was achieved. In this communication, we are discussing (1) to which extend the quality gain of the laboratory-based absorption-contrast tomography can be driven with respect to optical microscopy of stained tissue sections and (2) what value such a technique would add. As a proof of principle, four histological sections were affine-registered to corresponding three-dimensional (3D) tomography dataset. We are discussing a semi-automatic landmark-based 2D-3D registration framework and compare registration results based on mean square difference (MSD) metrics.

Applied physics letters 114(8), 083702 (2019). doi:10.1063/1.5085302, Advances in high-resolution hard X-ray computed tomography have led to the field of virtual histology to complement histopathological analyses. Phase-contrast modalities have been favored because, for soft tissues, the real part of the refractive index is orders of magnitude greater than the imaginary part. Nevertheless, absorption-contrast measurements of paraffin-embedded tissues have provided exceptionally high contrast combined with a submicron resolution. In this work, we present a quantitative comparison of phase tomography using synchrotron radiation-based X-ray double grating interferometry and conventional synchrotron radiation-based computed tomography in the context of histopathologically relevant paraffin-embedded human brain tissue. We determine the complex refractive index and compare the contrast-to-noise ratio (CNR) of each modality, accounting for the spatial resolution and optimizing the photon energy for absorption tomography. We demonstrate that the CNR in the phase modality is 1.6 times higher than the photon-energy optimized and spatial resolution-matched absorption measurements. We predict, however, that a further optimized phase tomography will provide a CNR gain of 4. This study seeks to boost the discussion of the relative merits of phase and absorption modalities in the context of paraffin-embedded tissues for virtual histology, highlighting the importance of optimization procedures for the two complementary modes and the trade-off between spatial and density resolution, not to mention the disparity in data acquisition and processing., Published by American Inst. of Physics, Melville, NY

X-ray imaging in absorption contrast mode is well established for hard tissue visualization. However, performance for lower density materials is limited due to a reduced contrast. Our aim is three-dimensional (3D) characterization of micro-morphology of human brain tissues down to (sub-)cellular resolution within a laboratory environment. Using the laboratory-based microtomography (μCT) system nanotom m (GE Sensing and Inspection Technologies GmbH, Wunstorf, Germany) and synchrotron radiation at the Diamond-Manchester Imaging Branchline I13-2 (Diamond Light Source, Didcot, UK), we have acquired 3D data with a resolution down to 0.45 μm for visualization of a human cerebellum specimen down to cellular level. We have shown that all selected modalities, namely laboratory-based absorption contrast micro-tomography (LBμCT), synchrotron radiation based in-line single distance phase contrast tomography (SDPR) and synchrotron radiation based single-grating interferometry (GI), can reach cellular resolution for tissue samples with a size in the mm-range. The results are discussed qualitatively in comparison to optical microscopy of haematoxylin and eosin (HE) stained sections. As phase contrast yields to a better data quality for soft tissues and in order to overcome restrictions of limited beamline access for phase contrast measurements, we have equipped the μCT system nanotom m with a double-grating phase contrast set-up. Preliminary experimental results of a knee sample consisting of a bony part and a cartilage demonstrate that phase contrast data exhibits better quality compared to absorption contrast. Currently, the set-up is under adjustment. It is expected that cellular resolution would also be achieved. The questions arise (1) what would be the quality gain of laboratory-based phase contrast in comparison to laboratory-based absorption contrast tomography and (2) could laboratory-based phase contrast data provide comparable results to synchrotron radiation based phase contrast data.

[Proceedings] - , 2016. - ISBN - doi:10.1117/12.2238702 Developments in X-Ray Tomography X, San Diego, California, 28 Aug 2016 - 1 Sep 2016; Proceedings of SPIE 9967, 99670O (2016). doi:10.1117/12.2238702, Heart disease is the number one cause of death worldwide. To improve therapy and patient outcome, the knowledge of anatomical changes in terms of lumen morphology and tissue composition of constricted arteries is crucial for designing a localized drug delivery to treat atherosclerosis disease. Traditional tissue characterization by histology is a pivotal tool, although it brings disadvantages such as vessel morphology modification during decalcification and slicing. X-ray tomography in absorption and phase contrast modes yields a deep understanding in blood vessel anatomy in healthy and diseased stages: measurements in absorption mode make visible highly absorbing tissue components including cholesterol plaques, whereas phase contrast tomography gains better contrast of the soft tissue components such as vessel walls. Established synchrotron radiation-based micro-CT techniques ensure high performance in terms of 3D visualization of highly absorbing and soft tissues., Published by SPIE, Bellingham, Wash.

Cell visualization and counting plays a crucial role in biological and medical research including the study of neurodegenerative diseases. The neuronal cell loss is typically determined to measure the extent of the disease. Its characterization is challenging because the cell density and size already differs by more than three orders of magnitude in a healthy cerebellum. Cell visualization is commonly performed by histology and fluorescence microscopy. These techniques are limited to resolve complex microstructures in the third dimension. Phase- contrast tomography has been proven to provide sufficient contrast in the three-dimensional imaging of soft tissue down to the cell level and, therefore, offers the basis for the three-dimensional segmentation. Within this context, a human cerebellum sample was embedded in paraffin and measured in local phase-contrast mode at the beamline ID19 (ESRF, Grenoble, France) and the Diamond Manchester Imaging Branchline I13-2 (Diamond Light Source, Didcot, UK). After the application of Frangi-based filtering the data showed sufficient contrast to automatically identify the Purkinje cells and to quantify their density to 177 cells per mm3 within the volume of interest. Moreover, brain layers were segmented in a region of interest based on edge detection. Subsequently performed histological analysis validated the presence of the cells, which required a mapping from the two- dimensional histological slices to the three-dimensional tomogram. The methodology can also be applied to further tissue types and shows potential for the computational tissue analysis in health and disease.

Micro computed tomography (mCT) is well established in virtually all fields of biomedical research, allowing for the non-destructive volumetric visualization of tissue morphology. A variety of specimens can be investigated, ranging from soft to hard tissue to engineered structures like scaffolds. Similarly, the size of the objects of interest ranges from a fraction of a millimeter to several tens of centimeters. While synchrotron radiation-based μCT still offers unrivaled data quality, the ever-improving technology of cathodic tube-based machines offers a valuable and more accessible alternative. The Biomaterials Science Center of the University of Basel operates a nanotomOR m (phoenix|x-ray, GE Sensing and Inspection Technologies GmbH, Wunstorf, Germany), with a 180 kV source and a minimal spot size of about 0.9 μm. Through the adjustable focus-specimen and focus-detector distances, the effective pixel size can be adjusted from below 500 nm to about 80 μm. On the high-resolution side, it is for example possible to visualize the tubular network in sub-millimeter thin dentin specimens. It is then possible to locally extract parameters such as tubule diameter, density, or alignment, giving information on cell movements during tooth formation. On the other side, with a horizontal shift of the 3,072 pixels x 2,400 pixels detector, specimens up to 35 cm in diameter can be scanned. It is possible, for example, to scan an entire human knee, albeit with inferior resolution. Lab source μCT machines are thus a powerful and flexible tool for the advancement of biomedical research, and a valuable and more accessible alternative to synchrotron radiation facilities.

X-ray imaging in the absorption contrast mode is an established method of visualising calcified tissues such as bone and teeth. Physically soft tissues such as brain or muscle are often imaged using magnetic resonance imaging (MRI). However, the spatial resolution of MRI is insufficient for identifying individual biological cells within three-dimensional tissue. X-ray grating interferometry (XGI) has advantages for the investigation of soft tissues or the simultaneous three-dimensional visualisation of soft and hard tissues. Since laboratory microtomography (μCT) systems have better accessibility than tomography set-ups at synchrotron radiation facilities, a great deal of effort has been invested in optimising XGI set-ups for conventional μCT systems. In this conference proceeding, we present how a two-grating interferometer is incorporated into a commercially available nanotom m (GE Sensing and Inspection Technologies GmbH) μCT system to extend its capabilities toward phase contrast. We intend to demonstrate superior contrast in spiders (Hogna radiata (Fam. Lycosidae) and Xysticus erraticus (Fam. Thomisidae)), as well as the simultaneous visualisation of hard and soft tissues. XGI is an imaging modality that provides quantitative data, and visualisation is an important part of biomimetics; consequently, hard X-ray imaging provides a sound basis for bioinspiration, bioreplication and biomimetics and allows for the quantitative comparison of biofabricated products with their natural counterparts.

X-ray grating interferometry-based computed tomography is a phase contrast imaging technique that provides non-destructive, quantitative, and three-dimensional visualization with contrast superior to traditional absorption-based techniques, especially for materials primarily composed of low Z elements, such as biological tissues. However, it relies on measurements of the lateral shift of an interference pattern and is thus susceptible to so-called phase wrapping artifacts, which mainly occur at the sample-air interface. In this work, we present an algorithm for removal of such artifacts in the case of cylindrical samples and an experiment to verify its accuracy. The proposed algorithm is applied to the sinogram after phase retrieval and prior to reconstruction by finding sample edges with the absorption sinogram and replacing regions of the phase wrapped sinogram with modeled data. Our measurements show that the algorithm removes artifacts and produces more accurate δ values, as validated by measurements without phase wrapping. Our correction algorithm allows for measurements without submerging the sample in a water bath, simplifying the experimental setup and avoiding motion artifacts from gas bubbles.

There is a lack of the necessary methodology for three-dimensional (3D) investigation of soft tissues with cellular resolution without staining or tissue transformation. Synchrotron radiation based hard X-ray in-line phase contrast tomography using single-distance phase reconstruction (SDPR) provides high spatial resolution and density contrast for the visualization of individual cells using a standard specimen preparation and data reconstruction. In this study, we demonstrate the 3D characterization of a formalin-fixed paraffin-embedded (FFPE) human cerebellum specimen by SDPR at the Diamond-Manchester Imaging Branchline I13-2 (Diamond Light Source, UK) at pixel sizes down to 0.45 μm. The approach enables visualization of cerebellar layers (Stratum moleculare and Stratum granulosum), the 3D characterization of individual cells (Purkinje, stellate and granule cells) and can even resolve some subcellular structures (nucleus and nucleolus of Purkinje cells). The tomographic results are qualitatively compared to hematoxylin and eosin (H&E) stained histological sections. We demonstrate the potential benefits of hard X-ray microtomography for the investigations of biological tissues in comparison to conventional histology.

In a dental office, every day X rays of teeth within the oral cavity are obtained. Caries induces a mineral loss and, therefore, becomes visible by reduced X-ray absorption. The detailed spatial distribution of the mineral loss, however, is inaccessible in conventional dental radiology, since the dose for such studies is intolerable. As a consequence, such measurements can only be performed after tooth extraction. We have taken advantage of synchrotron radiation-based micro computed tomography to characterize a human tooth with a rather small, natural caries lesion and an artificially induced lesion provoked by acidic etching. Both halves of the tooth were separately visualized from 2400 radiographs recorded at the beam line P07 / PETRA III (HASYLAB at DESY, Hamburg, Germany) with an asymmetric rotation axis at photon energy of 45 keV. Because of the setup, one finds an energy shift in the horizontal plane, to be corrected. After the appropriate three-dimensional registration of the data with the ones of the same crown using the better accessible phoenix nanotom® m of General Electric, Wunstorf, Germany, one can determine the joint histogram, which enable to calibrate the system with the conventional X-ray source.

Cardiovascular diseases are the number one cause of death and morbidity in the world. Understanding disease development in terms of lumen morphology and tissue composition of constricted arteries is essential to improve treatment and patient outcome. X-ray tomography provides non-destructive three-dimensional data with micrometer-resolution. However, a common problem is simultaneous visualization of soft and hard tissue-containing specimens, such as atherosclerotic human coronary arteries. Unlike absorption based techniques, where X-ray absorption strongly depends on atomic number and tissue density, phase contrast methods such as grating interferometry have significant advantages as the phase shift is only a linear function of the atomic number. We demonstrate that grating interferometry-based phase tomography is a powerful method to three-dimensionally visualize a variety of anatomical features in atherosclerotic human coronary arteries, including plaque, muscle, fat, and connective tissue. Three formalin-fixed, human coronary arteries were measured using advanced laboratory μCT. While this technique gives information about plaque morphology, it is impossible to extract the lumen morphology. Therefore, selected regions were measured using grating based phase tomography, sinograms were treated with a wavelet-Fourier filter to remove ring artifacts, and reconstructed data were processed to allow extraction of vessel lumen morphology. Phase tomography data in combination with conventional laboratory μCT data of the same specimen shows potential, through use of a joint histogram, to identify more tissue types than either technique alone. Such phase tomography data was also rigidly registered to subsequently decalcified arteries that were histologically sectioned, although the quality of registration was insufficient for joint histogram analysis.

The geometric characterization of low-voltage dielectric electro-active polymer (EAP) structures, comprised of nanometer thickness but areas of square centimeters, for applications such as artificial sphincters requires methods with nanometer precision. Direct optical detection is usually restricted to sub-micrometer resolution because of the wavelength of the light applied. Therefore, we propose to take advantage of the cantilever bending system with optical readout revealing a sub-micrometer resolution at the deflection of the free end. It is demonstrated that this approach allows us to detect bending of rather conventional planar asymmetric, dielectric EAP-structures applying voltages well below 10 V. For this purpose, we built 100 μm-thin silicone films between 50 nm-thin silver layers on a 25 μm-thin polyetheretherketone (PEEK) substrate. The increase of the applied voltage in steps of 50 V until 1 kV resulted in a cantilever bending that exhibits only in restricted ranges the expected square dependence. The mean laser beam displacement on the detector corresponded to 6 nm per volt. The apparatus will therefore become a powerful mean to analyze and thereby improve low-voltage dielectric EAP-structures to realize nanometer-thin layers for stack actuators to be incorporated into artificial sphincter systems for treating severe urinary and fecal incontinence.

Among the clinically relevant imaging techniques, computed tomography (CT) reaches the best spatial resolution. Sub-millimeter voxel sizes are regularly obtained. For investigations on true micrometer level lab-based μCT has become gold standard. The aim of the present study is the hierarchical investigation of a human knee post mortem using hard X-ray μCT. After the visualization of the entire knee using a clinical CT with a spatial resolution on the sub-millimeter range, a hierarchical imaging study was performed using a laboratory μCT system nanotom m. Due to the size of the whole knee the pixel length could not be reduced below 65 μm. These first two data sets were directly compared after a rigid registration using a cross-correlation algorithm. The μCT data set allowed an investigation of the trabecular structures of the bones. The further reduction of the pixel length down to 25 μm could be achieved by removing the skin and soft tissues and measuring the tibia and the femur separately. True micrometer resolution could be achieved after extracting cylinders of several millimeters diameters from the two bones. The high resolution scans revealed the mineralized cartilage zone including the tide mark line as well as individual calcified chondrocytes. The visualization of soft tissues including cartilage, was arranged by X-ray grating interferometry (XGI) at ESRF and Diamond Light Source. Whereas the high-energy measurements at ESRF allowed the simultaneous visualization of soft and hard tissues, the low-energy results from Diamond Light Source made individual chondrocytes within the cartilage visual.

Recently a novel grating based x-ray imaging approach called directional x-ray dark-field imaging was introduced. Directional x-ray dark-field imaging yields information about the local texture of structures smaller than the pixel size of the imaging system. In this work we extend the theoretical description and data processing schemes for directional dark-field imaging to strongly scattering systems, which could not be described previously. We develop a simple scattering model to account for these recent observations and subsequently demonstrate the model using experimental data. The experimental data includes directional dark-field images of polypropylene fibers and a human tooth slice.

During the last decade, several tissues and biomaterials for medical applications in replacing bony tissues have been developed. Three-dimensional cell clusters and mandibular bone grafts are two distinct examples of these developments. The characterization of the complex three-dimensional structures, however, is still mainly restricted on the twodimensional analysis of histological slices. The present paper examines the quantitative analysis of mandibular bone grafts and three-dimensional cell clusters on the basis of synchrotron radiation-based micro computed tomography measurements. An automated search of pre-defined microstructures through component labeling is applied to the real datasets in order to identify features that reside independently from other components. The examples demonstrate three levels of complexity: rather large pieces of bone augmentation material that touch each other, individual adipocytes difficult to automatically segment in a wet cluster and osmium-stained adipocyte exhibiting higher X-ray absorption than the surrounding tissue. Although the structures of interest such as the cells can be labeled, de-clustering of the components requires the incorporation of erosion and dilation algorithms.

In this paper we describe the design of different X-ray Talbot interferometers that have been built at the tomography beamline ID19 of the European Synchrotron Radiation Facility (ESRF) in Grenoble, France, and give a short review of performance characteristics, of current developments, and of the results obtained with these instruments so far. Among the applications so far, soft-tissue imaging has been a particular focus, as demonstrated in a recent paper by Schulz et al. (J. Roy. Soc. Interface, in press).

Direct composite fillings belong to widespread tooth restoration techniques in dental medicine. The procedure consists of successive steps, which include etching of the prepared tooth surface, bonding and placement of composite in incrementally built up layers. Durability and lifespan of the composite inlays strongly depend on the accurate completion of the individual steps to be also realized by students in dental medicine. Improper handling or nonconformity in the bonding procedure often lead to air enclos ures (bubbles) as well as to significant ga ps between the composite layers or at the margins of the restoration. Traditionally one analyzes the quality of the restoration cutting the tooth in an arbitrarily selected plane and inspecting this plane by conventional optical microscopy. Although the precision of this established method is satisfactory, it is restricted to the selected two-dimensional plane. Rather simple micro computed tomography (µCT) systems, such as SkyScan 1174

Cone-beam computed tomography, also termed digital volume tomography, has become a standard technique in dentistry, allowing for fast 3D jaw imaging including denture at moderate spatial resolution. More detailed X-ray images of restricted volumes for post-mortem studies in dental anthropology are obtained by means of micro computed tomography. The present study evaluates the impact of the pipe smoking wear on teeth morphology comparing the abraded tooth with its contra-lateral counterpart. A set of 60 teeth, loose or anchored in the jaw, from 12 dentitions have been analyzed. After the two contra-lateral teeth were scanned, one dataset has been mirrored before the two datasets were registered using affine and rigid registration algorithms. Rigid registration provides three translational and three rotational parameters to maximize the overlap of two rigid bodies. For the affine registration, three scaling factors are incorporated. Within the present investigation, affine and rigid registrations yield comparable values. The restriction to the six parameters of the rigid registration is not a limitation. The differences in size and shape between the tooth and its contra-lateral counterpart generally exhibit only a few percent in the non-abraded volume, validating that the contralateral tooth is a reasonable approximation to quantify, for example, the volume loss as the result of long-term clay pipe smoking. Therefore, this approach allows quantifying the impact of the pipe abrasion on the internal tooth morphology including root canal, dentin, and enamel volumes.

Minimally invasive deep brain neurosurgical interventions require a profound knowledge of the morphology of the human brain. Generic brain atlases are based on histology including multiple preparation steps during the sectioning and staining. In order to correct the distortions induced in the anisotropic, inhomogeneous soft matter and therefore improve the accuracy of brain atlases, a non-destructive 3D imaging technique with the required spatial and density resolution is of great significance. Micro computed tomography provides true micrometer resolution. The application to post mortem human brain, however, is questionable because the differences of the components concerning X-ray absorption are weak. Therefore, magnetic resonance tomography has become the method of choice for three-dimensional imaging of human brain. Because the spatial resolution of this method is limited, an alternative has to be found for the three-dimensional imaging of cellular microstructures within the brain. Therefore, the present study relies on the synchrotron radiationbased micro computed tomography in the recently developed grating-based phase contrast mode. Using data acquired at the beamline ID 19 (ESRF, Grenoble, France) we demonstrate that grating-based tomography yields premium images of human thalamus, which can be used for the correction of histological distortions by 3D non-rigid registration.

Synchrotron radiation-based micro computed tomography (SRµCT) is well established to determine the degree of mineralization in bony tissue. The present study demonstrates that the method can be likewise used for three-dimensional analyses of dentin de- and remineralization. Four dentin discs about 4 mm in diameter and 0.8 mm thick were prepared from freshly extracted human third molars. In order to study the de- and re-mineralization, three of them were treated with 10% citric acid for the period of 10 min. Nano-particulate bioactive glass made of SiO2, P2O5, CaO, Na2O served for the re-mineralization in physiological saline. This process was carried out at the incubation temperature of 37 °C for 1 and 7 d, respectively. The native and the treated discs were comparatively examined by SRµCT in absorption contrast mode. Already the visual inspection of the tomograms obtained reveals remarkable differences related to the mean X-ray absorption and internal microstructure. The de-mineralization led to a surface morphology characteristic for the treated dentin collagen matrix. The re-mineralized discs show a dependence on the period of the treatment with the bio-active glass suspension. Initial signs of the remineralization were clearly present already after 24 h of incubation. The disc incubated for 7 d exhibits a degree of mineralization comparable to the native control disc. Thus, SRµCT is a powerful non-destructive technique for the analysis of dentin de- and re-mineralization.

Optical measurements reveal the preferential orientation of nanostructures within polymer films, which results from the fabrication process including mechanical and thermal treatments. As the wavelength of the incident light is generally much larger than the characteristic dimensions of the molecular arrangement in semi-crystalline or amorphous polymers, the optical signal originates not directly from the nanostructure of the polymers. Linear dichroism measurements were correlated with synchrotron radiation-based x-ray scattering data on commercially available polyetheretherketone (PEEK) thin films (12 to 50 μm). Annealing changed the structure of amorphous films to semi-crystalline ones associated with the measured linear dichroism. The intensity of the measured anisotropic signal depended on the film thickness. While for wavelengths between 450 and 1100 nm the transmission was higher when the polarizer was parallel to the machine direction, for larger wavelengths maximum transmission was observed with the polarizer perpendicular to the machine direction indicating excitations parallel and perpendicular to the PEEK molecule axis, respectively. Annealing PEEK films at temperatures between 160 and 240°C decreased the transmission at 540 nm by a factor of two, whereas the anisotropy remained constant. x-ray scattering revealed strongest anisotropy for a periodicity of 15 nm in the machine direction of the cast film extrusion process. The long-range order of amorphous and semi-crystalline entities can explain the x-ray scattering data and the related optical anisotropy of casted PEEK films.