A. Cedola

Non-destructive determination of local strain with 100-nanometre spatial resolution

Structure sizes of ∼180 nm are now standard in microelectronics, and state-of-the-art fabrication techniques can reduce these to just a few tens of nanometres (ref. 1). But at these length scales, the strain induced at interfaces can locally distort the crystal lattice, which may in turn affect device performance in an unpredictable way. A means of non-destructively characterizing such strain fields with high spatial resolution and sensitivity is therefore highly desirable. One approach is to use Raman spectroscopy, but this is limited by the intrinsic ∼0.5-µm resolution limit of visible light probes. Techniques based on electron-beam diffraction can achieve the desired nanometre-scale resolution. But either they require complex sample preparation procedures (which may alter the original strain field) or they are sensitive to distortional (but not dilational) strain within only the top few tens of nanometres of the sample surface. X-rays, on the other hand, have a much greater penetration depth, but have not hitherto achieved strain analysis with sub-micrometre resolution. Here we describe a magnifying diffraction imaging procedure for X-rays which achieves a spatial resolution of 100 nm in one dimension and a sensitivity of 10-4 for relative lattice variations. We demonstrate the suitability of this procedure for strain analysis by measuring the strain depth profiles beneath oxidized lines on silicon crystals.

Phase contrast hard x-ray microscopy with submicron resolution

In this letter we present a hard x-ray phase contrast microscope based on the divergent and coherent beam exiting an x-ray waveguide. It uses lensless geometrical projection to magnify spatial variations in optical path length more than 700 times. Images of a nylon fiber and a gold test pattern were obtained with a resolution of 0.14 μm in one direction. Exposure times as short as 0.1 s gave already visible contrast, opening the way to high resolution, real time studies.

Properties of a submicrometer x‐ray beam at the exit of a waveguide

This report discusses the properties of a 13‐keV submicrometer x‐ray beam exiting from a waveguide. Waveguides for this spectral regime can be constructed by enclosing a low‐absorbing material between highly absorbant metals. Best performance is found for about 0.1 μm guiding layer thickness. Measurements of the photon beam size close to the exit and of the intensity distribution far from the exit will be presented. From these data one derives a beam size at the exit which is identical to the guiding layer thickness. This number being in the submicrometer range offers interesting perspectives for microscopy experiments in the hard x‐ray range.

High gain beam compression in new-generation thin-film x-ray waveguides

X-ray waveguides can compress an incident beam for microscopy applications above 8 keV photon energy to sizes smaller than 100 nm in one dimension, a range which is not routinely accessed with other x-ray optics (e.g., Fresnel zone plates). Beryllium, because of its low absorption, is expected to provide the highest intensity gain. Measured gains for a beryllium waveguide of 74 nm thickness exceed values of 100 at 13 and 20 keV photon energy, which is an improvement by an order of magnitude compared to previously reported performances. The same object works also at 8 keV with gain 43.

SUBMICROMETER X-RAY BEAM PRODUCTION BY A THIN FILM WAVEGUIDE

A hard x‐ray waveguide capable of reducing a 13 keV monochromatic beam in one direction from 30 to only 0.1 μm has been characterized with synchrotron radiation. The guiding structure consisted of a carbon layer sandwiched between two Ni layers sputter‐deposited onto float glass. A beam exiting at the end of the guide in guiding direction was observed when a resonance effect due to the formation of x‐ray standing waves in the total reflection region takes place. The measured beam divergence (1.3 mrad) agrees with expectations. The total efficiency (exiting photon flux/incident photon flux) can ideally exceed 10−2 but is only 10−4 for this first prototype.

Simultaneous submicrometric 3D imaging of the micro-vascular network and the neuronal system in a mouse spinal cord.

Faults in vascular (VN) and neuronal networks of spinal cord are responsible for serious neurodegenerative pathologies. Because of inadequate investigation tools, the lacking knowledge of the complete fine structure of VN and neuronal system represents a crucial problem. Conventional 2D imaging yields incomplete spatial coverage leading to possible data misinterpretation, whereas standard 3D computed tomography imaging achieves insufficient resolution and contrast. We show that X-ray high-resolution phase-contrast tomography allows the simultaneous visualization of three-dimensional VN and neuronal systems of ex-vivo mouse spinal cord at scales spanning from millimeters to hundreds of nanometers, with nor contrast agent nor sectioning and neither destructive sample-preparation. We image both the 3D distribution of micro-capillary network and the micrometric nerve fibers, axon-bundles and neuron soma. Our approach is very suitable for pre-clinical investigation of neurodegenerative pathologies and spinal-cord-injuries, in particular to resolve the entangled relationship between VN and neuronal system.

A three-image algorithm for hard x-ray grating interferometry

A three-image method to extract absorption, refraction and scattering information for hard x-ray grating interferometry is presented. The method comprises a post-processing approach alternative to the conventional phase stepping procedure and is inspired by a similar three-image technique developed for analyzer-based x-ray imaging. Results obtained with this algorithm are quantitatively comparable with phase-stepping. This method can be further extended to samples with negligible scattering, where only two images are needed to separate absorption and refraction signal. Thanks to the limited number of images required, this technique is a viable route to bio-compatible imaging with x-ray grating interferometer. In addition our method elucidates and strengthens the formal and practical analogies between grating interferometry and the (non-interferometric) diffraction enhanced imaging technique.

Debye function analysis and 2D imaging of nanoscaled engineered bone

The Debye Function Analysis of diffraction patterns from nanosized mineral crystals showing different average degrees of maturity was carried out on engineered bone samples. The analysis relied on a bivariate family of atomistic hydroxyapatite nanocrystal models and provided information about crystal structure, size and shape distributions of the mineral component of the newly formed bone. An average rod-like shape of nanocrystals was found in all samples, with average sizes well matching the collagen I gap region. The diffraction patterns investigated through the Debye Function Analysis were used as signal models to perform the Canonical Correlation Analysis of high resolution X-ray micro-diffraction patterns collected on porous and resorbable hydroxyapatite/silicon-stabilized tricalcium phosphate (Si-TCP) implants. The nosologic maps clearly showed a size gradient in the new formed bone that validates the mechanism (mimicking the bone remodelling in orthotopic bones) of a continuous deposition of bone by osteoblasts, an increasing mineralization of the newly deposited bone, a growth of the new crystals, at the same time that osteoclasts adhere to the scaffold surface and resorb the bioceramic. The comparison of samples at different implantation times proved that the selective resorption of Si-TCP component from the scaffold was already evident after two and almost complete after six months.

Engineered bone from bone marrow stromal cells : a structural study by an advanced x-ray microdiffraction technique

The mechanism of mineralized matrix deposition was studied in a tissue engineering approach in which bone tissue is formed when porous ceramic constructs are loaded with bone marrow stromal cells and implanted in vivo. We investigated the local interaction between the mineral crystals of the engineered bone and the biomaterial by means of microdiffraction, using a set-up based on an x-ray waveguide. We demonstrated that the newly formed bone is well organized inside the scaffold pore, following the growth model of natural bone. Combining wide angle (WAXS) and small angle (SAXS) x-ray scattering with high spatial resolution, we were able to determine the orientation of the crystallographic c-axis inside the bone crystals, and the orientation of the mineral crystals and collagen micro-fibrils with respect to the scaffold. In this work we analysed six samples and for each of them two pores were studied in detail. Similar results were obtained in all cases but we report here only the most significant sample.

New insights on the biomineralisation process developing in human lungs around inhaled asbestos fibres

Once penetrated into the lungs of exposed people, asbestos induces an in vivo biomineralisation process that leads to the formation of a ferruginous coating embedding the fibres. The ensemble of the fibre and the coating is referred to as asbestos body and is believed to be responsible for the high toxicological outcome of asbestos. Lung tissue of two individuals subjected to prolonged occupational exposure to crocidolite asbestos was investigated using synchrotron radiation micro-probe tools. The distribution of K and of elements heavier than Fe (Zn, Cu, As, and Ba) in the asbestos bodies was observed for the first time. Elemental quantification, also reported for the first time, confirmed that the coating is highly enriched in Fe (~20% w/w), and x-ray absorption spectroscopy indicated that Fe is in the 3+ oxidation state and that it is present in the form of ferritin or hemosiderin. Comparison of the results obtained studying the asbestos bodies upon removing the biological tissue by chemical digestion and those embedded in histological sections, allowed unambiguously distinguishing the composition of the asbestos bodies, and understanding to what extent the digestion procedure altered their chemical composition. A speculative model is proposed to explain the observed distribution of Fe.