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An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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Abdominal aortic and popliteal artery aneurysms are vascular diseases which show massive degeneration, weakening of the vascular wall and loss of the vascular tissue functionality. They are driven by inflammatory, hemodynamical factors and biological alterations that may lead, in the case of an abdominal aortic aneurysm, to sudden and dangerous ruptures of the arteries. Here, human aortic and popliteal aneurysm tissues were obtained during surgical repair, and studied by synchrotron radiation X-ray scanning microdiffraction and small-angle scattering, to investigate the microcalcifications present in the tissues. Data collected during the experiments were transformed into quantitative microscopy images through the combination of statistical approaches and crystallographic methods. As a result of this multi-step analysis, microcalcifications, which are markers of the pathology, were classified in terms of chemical and structural content. This analysis helped to identify the presence of nanocrystalline hy-droxy-apatite and microcrystalline cholesterol, embedded in myofilament, and elastin-containing tissue with low collagen content in predominantly nanocrystalline areas. The generality of the approach allows it to be transferred to other types of tissue and other pathologies affected by microcalcifications, such as thyroid carcinoma, breast cancer, testicular microli-thia-sis or glioblastoma.
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In X-ray computed tomography, the task of imaging only a local region of interest (ROI) inside a larger sample is very important. However, without a priori information, this ROI cannot be exactly reconstructed using only the image data limited to the ROI. We propose here an approach of region-of-interest tomography, which reconstructs a ROI within an object from projections of different fields of view acquired on a specific angular sampling scheme in the same tomographic experiment. We present a stable procedure that not only yields high-quality images of the ROI but keeps as well the quantitative contrast on the reconstructed images. In addition, we analyze the minimum number of projections required for ROI tomography from the point of view of the band region of the Radon transform, which confirms this number must be estimated based on the size of the entire object and not only on the size of the ROI.
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Procesamiento de Imagen Asistido por Computador/métodos , Fantasmas de Imagen , Tomografía Computarizada por Rayos X/métodos , Algoritmos , Simulación por Computador , HumanosRESUMEN
For many scientific questions gaining three-dimensional insight into a specimen can provide valuable information. We here present an instrument called "tOMography Nano crYo (OMNY)," dedicated to high resolution 3D scanning x-ray microscopy at cryogenic conditions via hard X-ray ptychography. Ptychography is a lens-less imaging method requiring accurate sample positioning. In OMNY, this in achieved via dedicated laser interferometry and closed-loop position control reaching sub-10 nm positioning accuracy. Cryogenic sample conditions are maintained via conductive cooling. 90 K can be reached when using liquid nitrogen as coolant, and 10 K is possible with liquid helium. A cryogenic sample-change mechanism permits measurements of cryogenically fixed specimens. We compare images obtained with OMNY with older measurements performed using a nitrogen gas cryo-jet of stained, epoxy-embedded retina tissue and of frozen-hydrated Chlamydomonas cells.
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Imagenología Tridimensional/instrumentación , Microscopía/instrumentación , Tomografía por Rayos X/instrumentación , Animales , Chlamydomonas , Criopreservación/instrumentación , Diseño de Equipo , Interferometría/instrumentación , Rayos Láser , Imagen Óptica/instrumentación , Retina/citología , Dispersión del Ángulo Pequeño , Temperatura , Difracción de Rayos X/instrumentaciónRESUMEN
Bovine cornea was studied with scanning small-angle X-ray scattering (SAXS) microscopy, by using both synchrotron radiation and a microfocus laboratory source. A combination of statistical (adaptive binning and canonical correlation analysis) and crystallographic (pair distribution function analysis) approaches allowed inspection of the collagen lateral packing of the supramolecular structure. Results reveal (i) a decrease of the interfibrillar distance and of the shell thickness around the fibrils from the periphery to the center of the cornea, (ii) a uniform fibril diameter across the explored area, and (iii) a distorted quasi-hexagonal arrangement of the collagen fibrils. The results are in agreement with existing literature. The overlap between laboratory and synchrotron-radiation data opens new perspectives for further studies on collagen-based/engineered tissues by the SAXS microscopy technique at laboratory-scale facilities.
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X-ray ptychography is a scanning variant of coherent diffractive imaging with the ability to image large fields of view at high resolution. It further allows imaging of non-isolated specimens and can produce quantitative mapping of the electron density distribution in 3D when combined with computed tomography. The method does not require imaging lenses, which makes it dose efficient and suitable to multi-keV X-rays, where efficient photon counting, pixelated detectors are available. Here we present the first highly resolved quantitative X-ray ptychographic tomography of an extended object yielding 16â nm isotropic 3D resolution recorded at 2 Å wavelength. This first-of-its-kind demonstration paves the way for ptychographic X-ray tomography to become a promising method for X-ray imaging of representative sample volumes at unmatched resolution, opening tremendous potential for characterizing samples in materials science and biology by filling the resolution gap between electron microscopy and other X-ray imaging techniques.
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We present a small-angle x-ray scattering study of crystals formed by temperature-sensitive, swollen microgel particles consisting of poly(N-isopropylacrylamide) copolymerized with acrylic acid and 5 mol % of a cross-linker. As for hard spheres, the random hexagonal close-packed structure is predominant during crystal growth and slowly transforms toward the face-centered-cubic structure. However, a transient phase of body-centered-cubic crystal is observed in an intermediate range of effective volume fractions. We estimate that the studied suspensions are close to a transition from face-centered-cubic to body-centered-cubic structure that can be understood by the tendency of the system to maximize the excluded volume and minimize the contact area between the particles.
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We present an instrument dedicated to 3D scanning x-ray microscopy, allowing a sample to be precisely scanned through a beam while the angle of x-ray incidence can be changed. The position of the sample is controlled with respect to the beam-defining optics by laser interferometry. The instrument achieves a position stability better than 10 nm standard deviation. The instrument performance is assessed using scanning x-ray diffraction microscopy and we demonstrate a resolution of 18 nm in 2D imaging of a lithographic test pattern while the beam was defined by a pinhole of 3 µm in diameter. In 3D on a test object of copper interconnects of a microprocessor, a resolution of 53 nm is achieved.
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Scanning small and wide angle X-ray scattering (scanning SWAXS) experiments were performed on healthy and pathologic human bone sections. Via crystallographic tools the data were transformed into quantitative images and as such compared with circularly polarized light (CPL) microscopy images. SWAXS and CPL images allowed extracting information of the mineral nanocrystalline phase embedded, with and without preferred orientation, in the collagen fibrils, mapping local changes at sub-osteon resolution. This favorable combination has been applied for the first time to biopsies of dwarfism syndrome and Paget's disease to shed light onto the cortical structure of natural bone in healthy and pathologic sections.
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Enfermedades Óseas/patología , Huesos/anatomía & histología , Luz , Microscopía Electrónica de Rastreo/métodos , Dispersión de Radiación , Adulto , Anciano de 80 o más Años , Huesos/ultraestructura , Cristalografía por Rayos X , Enanismo/patología , Osteón/anatomía & histología , Humanos , Masculino , Microscopía de Polarización/métodos , Osteítis Deformante/patología , Reproducibilidad de los Resultados , Dispersión del Ángulo Pequeño , Síndrome , Difracción de Rayos XRESUMEN
In this work we demonstrate the feasibility of applying small-angle X-ray scattering computed tomography (SAXS-CT) for non-invasive molecular imaging of myelin sheaths in a rat brain. Our results show that the approach yields information on several quantities, including the relative myelin concentration, its periodicity, the total thickness of the myelin sheaths, and the relative concentration of cytoskeletal neurofilaments. For example the periodicity of the myelin sheaths varied in the range from 17.0 to 18.2 nm around an average of 17.6 (±0.3) nm. We believe that imaging, i.e., spatially resolved measuring these quantities could provide general means for understanding the relation to a number of neurodegenerative diseases.
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Encéfalo/ultraestructura , Imagenología Tridimensional/métodos , Vaina de Mielina/ultraestructura , Dispersión del Ángulo Pequeño , Tomografía Computarizada por Rayos X/métodos , Difracción de Rayos X/métodos , Animales , Encéfalo/diagnóstico por imagen , Vaina de Mielina/diagnóstico por imagen , RatasRESUMEN
A growing number of X-ray sources based on the free-electron laser (XFEL) principle are presently under construction or have recently started operation. The intense, ultrashort pulses of these sources will enable new insights in many different fields of science. A key problem is to provide x-ray optical elements capable of collecting the largest possible fraction of the radiation and to focus into the smallest possible focus. As a key step towards this goal, we demonstrate here the first nanofocusing of hard XFEL pulses. We developed diamond based Fresnel zone plates capable of withstanding the full beam of the world's most powerful x-ray laser. Using an imprint technique, we measured the focal spot size, which was limited to 320 nm FWHM by the spectral band width of the source. A peak power density in the focal spot of 4×10(17)W/cm(2) was obtained at 70 fs pulse length.
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The basic principles of x-ray image formation in radiology have remained essentially unchanged since Röntgen first discovered x-rays over a hundred years ago. The conventional approach relies on x-ray attenuation as the sole source of contrast and draws exclusively on ray or geometrical optics to describe and interpret image formation. Phase-contrast or coherent scatter imaging techniques, which can be understood using wave optics rather than ray optics, offer ways to augment or complement the conventional approach by incorporating the wave-optical interaction of x-rays with the specimen. With a recently developed approach based on x-ray optical gratings, advanced phase-contrast and dark-field scatter imaging modalities are now in reach for routine medical imaging and non-destructive testing applications. To quantitatively assess the new potential of particularly the grating-based dark-field imaging modality, we here introduce a mathematical formalism together with a material-dependent parameter, the so-called linear diffusion coefficient and show that this description can yield quantitative dark-field computed tomography (QDFCT) images of experimental test phantoms.
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Oscuridad , Tomografía Computarizada por Rayos X/métodos , Difusión , Procesamiento de Imagen Asistido por Computador , Modelos Lineales , Fantasmas de ImagenRESUMEN
Scanning transmission X-ray microscopy (STXM) is a powerful imaging technique, in which a small X-ray probe is raster scanned across a specimen. Complete knowledge of the complex-valued transmission function of the specimen can be gained using detection schemes whose every-day use, however, is often hindered by the need of specialized configured detectors or by slow or noisy readout of area detectors. We report on sub-50 nm-resolution STXM studies in the hard X-ray regime using the PILATUS, a fully pixelated fast framing detector operated in single-photon counting mode. We demonstrate a range of imaging modes, including phase contrast and dark-field imaging.
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A synchrotron study is presented in which the concept of one-dimensional tomographic reconstruction of small-angle X-ray scattering patterns is applied to investigate polyamide 6 monofilaments, dip-coated with alumina particles. The filaments are scanned with a focused synchrotron beam and the resulting scattering patterns are recorded with a PILATUS 2M detector. The reconstructed sequence of SAXS images reflects the local nanostructure variation along the filament radius. In particular, the influence of coating process parameters on the polyamide 6 is investigated.
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Synovial joints articulate in a lubricating environment, the system providing for smooth articulation. The articular cartilage overlying the bone consists of a network of collagen fibres. This network is essential to cartilage integrity, suffering damage in degenerative joint disease such as osteoarthritis. At Surrey and also in work conducted by this group at the Paul Scherrer Institute (PSI) synchrotron site we have been applying a number of techniques to study the bone-cartilage interface and of changes occurring in this with disease. One of the techniques attracting particular interest is X-ray phase contrast imaging, yielding information on anatomical features that manifest from the large scale organisation of collagen and the mineralised phase contained within the collagen fibres in the deep cartilage zone. This work briefly reviews some of the basic supporting physics of X-ray phase contrast imaging and then shows example images of the articular surface and subchondral bone and other supporting results obtained to-date. Present results have been obtained on sections of bone not displaying evidence of an osteoarthritic lesion and can be used as a baseline against which diseased bone can be compared.
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Cartílago Articular/diagnóstico por imagen , Fémur/diagnóstico por imagen , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Tomografía Computarizada por Rayos X/métodos , Difracción de Rayos X/métodos , Animales , PorcinosRESUMEN
Micro Proton Induced X-ray Emission (micro-PIXE) analysis has been employed herein in investigating and quantifying the distribution of a number of essential elements in thin human diseased articular cartilage sections affected by osteoarthritis (OA). Various cations Ca, P and Zn have been reported to play an important role both in the normal growth and remodelling of articular cartilage and subchondral bone as well as in the degenerative and inflammatory processes associated with the disease; they act as co-factors of a class of enzymes known as metalloproteinases which are believed to be active during the initiation, progress and remodelling processes associated with osteoarthritis. Other important enzymes such as alkaline phosphatase are associated with cartilage mineralization. Synchrotron radiation X-ray fluorescence (SR-XRF) for mapping of elemental distributions in bone and cartilage has also been employed by the present group and others. In the current investigations using the cSAXS beamline at the Swiss light source, Small-Angle X-ray Scattering (SAXS) was carried out on decalcified human articular cartilage to explore the structural and organizational changes of collagen networks in diseased articular cartilage.
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Biomarcadores/análisis , Cartílago Articular/química , Fémur/química , Osteoartritis de la Cadera/metabolismo , Dispersión del Ángulo Pequeño , Espectrometría por Rayos X/métodos , Difracción de Rayos X/métodos , Humanos , Osteoartritis de la Cadera/diagnósticoRESUMEN
X-ray photon correlation spectroscopy (XPCS) provides an opportunity to study the dynamics of systems by measuring the temporal fluctuations in a far-field diffraction pattern. A two-dimensional detector system has been developed to investigate fluctuations in the frequency range of several Hz to kHz. The X-ray detector system consists of a thin 100 microm scintillation crystal coupled to a Geiger-mode avalanche photodiode array. In this article the elements of the system are detailed and the detector for XPCS measurements is demonstrated.
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We develop a novel method for structure determination of confined fluids using diffraction-grating-based x-ray interferometry.Within this approach, diffraction from a microfluidic array, which acts both as confinement and transmission diffraction grating, provides the reference wave, whereas the density modulations of the confined fluid, acting as a weak phase object, generate the object wave. The ensemble-averaged density profile of the fluid perpendicular to the confining channel is then unambiguously obtained from the interference between the reference and object waves by direct Fourier inversion.
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Mezclas Complejas/química , Diseño Asistido por Computadora , Interferometría/instrumentación , Microfluídica/instrumentación , Modelos Teóricos , Refractometría/instrumentación , Difracción de Rayos X/instrumentación , Simulación por Computador , Diseño de Equipo , Análisis de Falla de Equipo , Luz , Dispersión de RadiaciónRESUMEN
The influence of different physical parameters, such as the source size and the energy spectrum, on the functional capability of a grating interferometer applied for phase-contrast imaging is discussed using numerical simulations based on Fresnel diffraction theory. The presented simulation results explain why the interferometer could be well combined with polychromatic laboratory x-ray sources in recent experiments. Furthermore, it is shown that the distance between the two gratings of the interferometer is not in general limited by the width of the photon energy spectrum. This implies that interferometers that give a further improved image quality for phase measurements can be designed, because the primary measurement signal for phase measurements can be increased by enlargement of this distance. Finally, the mathematical background and practical instructions for the quantitative evaluation of measurement data acquired with a polychromatic x-ray source are given.
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We report numerical and experimental results demonstrating accurate region-of-interest computed tomography (CT) reconstruction based on differential phase-contrast projection (DPC) images. The approach removes the constraint of covering the entire sample within the field of view of the image detector. Particularly for biomedical applications, the presented DPC-CT region-of-interest approach will allow for the visualization of previously inaccessible details deep inside an entire animal or organ. We envisage that this development will also be of interest for potential future clinical applications, because grating-based DPC-CT can be implemented with standard x-ray tube sources.