High-resolution non-destructive three-dimensional imaging of integrated circuits.

Modern nanoelectronics has advanced to a point at which it is impossible to image entire devices and their interconnections non-destructively because of their small feature sizes and the complex three-dimensional structures resulting from their integration on a chip. This metrology gap implies a lack of direct feedback between design and manufacturing processes, and hampers quality control during production, shipment and use. Here we demonstrate that X-ray ptychography-a high-resolution coherent diffractive imaging technique-can create three-dimensional images of integrated circuits of known and unknown designs with a lateral resolution in all directions down to 14.6 nanometres. We obtained detailed device geometries and corresponding elemental maps, and show how the devices are integrated with each other to form the chip. Our experiments represent a major advance in chip inspection and reverse engineering over the traditional destructive electron microscopy and ion milling techniques. Foreseeable developments in X-ray sources, optics and detectors, as well as adoption of an instrument geometry optimized for planar rather than cylindrical samples, could lead to a thousand-fold increase in efficiency, with concomitant reductions in scan times and voxel sizes.

Fast and accurate data collection for macromolecular crystallography using the JUNGFRAU detector.

The accuracy of X-ray diffraction data is directly related to how the X-ray detector records photons. Here we describe the application of a direct-detection charge-integrating pixel-array detector (JUNGFRAU) in macromolecular crystallography (MX). JUNGFRAU features a uniform response on the subpixel level, linear behavior toward high photon rates, and low-noise performance across the whole dynamic range. We demonstrate that these features allow accurate MX data to be recorded at unprecedented speed. We also demonstrate improvements over previous-generation detectors in terms of data quality, using native single-wavelength anomalous diffraction (SAD) phasing, for thaumatin, lysozyme, and aminopeptidase N. Our results suggest that the JUNGFRAU detector will substantially improve the performance of synchrotron MX beamlines and equip them for future synchrotron light sources.

Nanostructure surveys of macroscopic specimens by small-angle scattering tensor tomography.

The mechanical properties of many materials are based on the macroscopic arrangement and orientation of their nanostructure. This nanostructure can be ordered over a range of length scales. In biology, the principle of hierarchical ordering is often used to maximize functionality, such as strength and robustness of the material, while minimizing weight and energy cost. Methods for nanoscale imaging provide direct visual access to the ultrastructure (nanoscale structure that is too small to be imaged using light microscopy), but the field of view is limited and does not easily allow a full correlative study of changes in the ultrastructure over a macroscopic sample. Other methods of probing ultrastructure ordering, such as small-angle scattering of X-rays or neutrons, can be applied to macroscopic samples; however, these scattering methods remain constrained to two-dimensional specimens or to isotropically oriented ultrastructures. These constraints limit the use of these methods for studying nanostructures with more complex orientation patterns, which are abundant in nature and materials science. Here, we introduce an imaging method that combines small-angle scattering with tensor tomography to probe nanoscale structures in three-dimensional macroscopic samples in a non-destructive way. We demonstrate the method by measuring the main orientation and the degree of orientation of nanoscale mineralized collagen fibrils in a human trabecula bone sample with a spatial resolution of 25 micrometres. Symmetries within the sample, such as the cylindrical symmetry commonly observed for mineralized collagen fibrils in bone, allow for tractable sampling requirements and numerical efficiency. Small-angle scattering tensor tomography is applicable to both biological and materials science specimens, and may be useful for understanding and characterizing smart or bio-inspired materials. Moreover, because the method is non-destructive, it is appropriate for in situ measurements and allows, for example, the role of ultrastructure in the mechanical response of a biological tissue or manufactured material to be studied.

LamNI - an instrument for X-ray scanning microscopy in laminography geometry.

Across all branches of science, medicine and engineering, high-resolution microscopy is required to understand functionality. Although optical methods have been developed to `defeat' the diffraction limit and produce 3D images, and electrons have proven ever more useful in creating pictures of small objects or thin sections, so far there is no substitute for X-ray microscopy in providing multiscale 3D images of objects with a single instrument and minimal labeling and preparation. A powerful technique proven to continuously access length scales from 10 nm to 10 µm is ptychographic X-ray computed tomography, which, on account of the orthogonality of the tomographic rotation axis to the illuminating beam, still has the limitation of necessitating pillar-shaped samples of small (ca 10 µm) diameter. Large-area planar samples are common in science and engineering, and it is therefore highly desirable to create an X-ray microscope that can examine such samples without the extraction of pillars. Computed laminography, where the axis of rotation is not perpendicular to the illumination direction, solves this problem. This entailed the development of a new instrument, LamNI, dedicated to high-resolution 3D scanning X-ray microscopy via hard X-ray ptychographic laminography. Scanning precision is achieved by a dedicated interferometry scheme and the instrument covers a scan range of 12 mm × 12 mm with a position stability of 2 nm and positioning errors below 5 nm. A new feature of LamNI is a pair of counter-rotating stages carrying the sample and interferometric mirrors, respectively.

Silicon carbide X-ray beam position monitors for synchrotron applications.

In this work, the performance of thin silicon carbide membranes as material for radiation hard X-ray beam position monitors (XBPMs) is investigated. Thermal and electrical behavior of XBPMs made from thin silicon carbide membranes and single-crystal diamond is compared using finite-element simulations. Fabricated silicon carbide devices are also compared with a 12 µm commercial polycrystalline diamond XBPM at the Swiss Light Source at the Paul Scherrer Institute. Results show that silicon carbide devices can reach equivalent transparencies while showing improved linearity, dynamics and signal-to-noise ratio compared with commercial polycrystalline diamond XBPMs. Given the obtained results and availability of electronic-grade epitaxies on up to 6 inch wafers, it is expected that silicon carbide can substitute for diamond in most beam monitoring applications, whereas diamond, owing to its lower absorption, could remain the material of choice in cases of extreme X-ray power densities, such as pink and white beams.

Ptychographic X-ray computed tomography at the nanoscale.

X-ray tomography is an invaluable tool in biomedical imaging. It can deliver the three-dimensional internal structure of entire organisms as well as that of single cells, and even gives access to quantitative information, crucially important both for medical applications and for basic research. Most frequently such information is based on X-ray attenuation. Phase contrast is sometimes used for improved visibility but remains significantly harder to quantify. Here we describe an X-ray computed tomography technique that generates quantitative high-contrast three-dimensional electron density maps from phase contrast information without reverting to assumptions of a weak phase object or negligible absorption. This method uses a ptychographic coherent imaging approach to record tomographic data sets, exploiting both the high penetration power of hard X-rays and the high sensitivity of lensless imaging. As an example, we present images of a bone sample in which structures on the 100 nm length scale such as the osteocyte lacunae and the interconnective canalicular network are clearly resolved. The recovered electron density map provides a contrast high enough to estimate nanoscale bone density variations of less than one per cent. We expect this high-resolution tomography technique to provide invaluable information for both the life and materials sciences.

Three-dimensional mass density mapping of cellular ultrastructure by ptychographic X-ray nanotomography.

We demonstrate absolute quantitative mass density mapping in three dimensions of frozen-hydrated biological matter with an isotropic resolution of 180 nm. As model for a biological system we use Chlamydomonas cells in buffer solution confined in a microcapillary. We use ptychographic X-ray computed tomography to image the entire specimen, including the 18 µm-diameter capillary, thereby providing directly an absolute mass density measurement of biological matter with an uncertainty of about 6%. The resulting maps have sufficient contrast to distinguish cells from the surrounding ice and several organelles of different densities inside the cells. Organelles are identified by comparison with a stained, resin-embedded specimen, which can be compared with established transmission electron microscopy results. For some identified organelles, the knowledge of their elemental composition reduces the uncertainty of their mass density measurement down to 1% with values consistent with previous measurements of dry weight concentrations in thin cellular sections by scanning transmission electron microscopy. With prospects of improving the spatial resolution in the near future, we expect that the capability of non-destructive three-dimensional mapping of mass density in biological samples close to their native state becomes a valuable method for measuring the packing of organic matter on the nanoscale.

Mass density and water content of saturated never-dried calcium silicate hydrates.

Calcium silicate hydrates (C-S-H) are the most abundant hydration products in ordinary Portland cement paste. Yet, despite the critical role they play in determining mechanical and transport properties, there is still a debate about their density and exact composition. Here, the site-specific mass density and composition of C-S-H in hydrated cement paste are determined with nanoscale resolution in a nondestructive approach. We used ptychographic X-ray computed tomography in order to determine spatially resolved mass density and water content of the C-S-H within the microstructure of the cement paste. Our findings indicate that the C-S-H at the border of hydrated alite particles possibly have a higher density than the apparent inner-product C-S-H, which is contrary to the common expectations from previous works on hydrated cement paste.

Coherent x-ray imaging of collagen fibril distributions within intact tendons.

The characterization of the structure of highly hierarchical biosamples such as collagen-based tissues at the scale of tens of nanometers is essential to correlate the tissue structure with its growth processes. Coherent x-ray Bragg ptychography is an innovative imaging technique that gives high resolution images of the ordered parts of such samples. Herein, we report how we used this method to image the collagen fibrillar ultrastructure of intact rat tail tendons. The images show ordered fibrils extending over 10-20 µm in length, with a quantifiable D-banding spacing variation of 0.2%. Occasional defects in the fibrils distribution have also been observed, likely indicating fibrillar fusion events.

High-throughput ptychography using Eiger: scanning X-ray nano-imaging of extended regions.

The smaller pixel size and high frame rate of next-generation photon counting pixel detectors opens new opportunities for the application of X-ray coherent diffractive imaging (CDI). In this manuscript we demonstrate fast image acquisition for ptychography using an Eiger detector. We achieve above 25,000 resolution elements per second, or an effective dwell time of 40 µs per resolution element, when imaging a 500 µm × 290 µm region of an integrated electronic circuit with 41 nm resolution. We further present the application of a scheme of sharing information between image parts that allows the field of view to exceed the range of the piezoelectric scanning system and requirements on the stability of the illumination to be relaxed.

Coherent X-ray Imaging: Bridging the Gap between Atomic and Micro-scale Investigations.

We present a review of state-of-the art X-ray imaging techniques based on partially coherent synchrotron radiation. Full-field X-ray tomography, X-ray ptychography, scanning small-angle X-ray scattering, and scanning transmission X-ray microscopy are imaging techniques that gather structural information at spatial resolution ranging from several microns to a few tens of nanometers in both real- and reciprocal space. These methods exploit contrast mechanisms based on absorption, phase, and spectroscopic signals. We provide examples of how these techniques can be applied to address scientific questions ranging from imaging of biological samples, to foam rheology, and cement composition.

Coherent X-ray imaging: bridging the gap between atomic and micro-scale investigations.

We present a review of state-of-the art X-ray imaging techniques based on partially coherent synchrotron radiation. Full-field X-ray tomography, X-ray ptychography, scanning small-angle X-ray scattering, and scanning transmission X-ray microscopy are imaging techniques that gather structural information at spatial resolution ranging from several microns to a few tens of nanometers in both real- and reciprocal space. These methods exploit contrast mechanisms based on absorption, phase, and spectroscopic signals. We provide examples of how these techniques can be applied to address scientific questions ranging from imaging of biological samples, to foam rheology, and cement composition.

Travelling through the Natural Hierarchies of Type I Collagen with X-rays: From Tendons of Cattle, Horses, Sheep and Pigs.

Type I collagen physiological scaffold for tissue regeneration is considered one of the widely used biomaterials for tissue engineering and medical applications. It is hierarchically organized: five laterally staggered molecules are packed within fibrils, arranged into fascicles and bundles. The structural organization is correlated to the direction and intensity of the forces which can be loaded onto the tissue. For a tissue-specific regeneration, the required macro- and microstructure of a suitable biomaterial has been largely investigated. Conversely, the function of multiscale structural integrity has been much less explored but is crucial for scaffold design and application. In this work, collagen was extracted from different animal sources with protocols that alter its structure. Collagen of tendon shreds excised from cattle, horse, sheep and pig was structurally investigated by wide- and small-angle X-ray scattering techniques, at both molecular and supramolecular scales, and thermo-mechanically with thermal and load-bearing tests. Tendons were selected because of their resistance to chemical degradation and mechanical stresses. The multiscale structural integrity of tendons' collagen was studied in relation to the animal source, anatomic location and source for collagen extraction.

Morphology of lamellae-forming block copolymer films between two orthogonal chemically nanopatterned striped surfaces.

The structure of block copolymers results from the interplay between weak intermolecular forces, typically in the order of k(B)T per molecule. This is particularly true for block copolymer thin films in the presence of chemically patterned surfaces, where the different contributions to the total free energy, the interfacial and bulklike terms, have comparable magnitudes. Here, we report on the structures formed by block copolymers films equilibrated between two chemically patterned surfaces with orthogonal stripes. Our experiments and simulations reveal that the domains are continuous through the film and the interface between domains resembles the Scherk's first minimal surface. The impact of chemical patterns on block copolymer morphologies and the underlying physics gives insight into the nanofabrication of complex nanostructures with directed self-assembly using two engineered boundary conditions, as opposed to only one.

Characterization of high-resolution diffractive X-ray optics by ptychographic coherent diffractive imaging.

We have employed ptychographic coherent diffractive imaging to completely characterize the focal spot wavefield and wavefront aberrations of a high-resolution diffractive X-ray lens. The ptychographic data from a strongly scattering object was acquired using the radiation cone emanating from a coherently illuminated Fresnel zone plate at a photon energy of 6.2 keV. Reconstructed images of the object were retrieved with a spatial resolution of 8 nm by combining the difference-map phase retrieval algorithm with a non-linear optimization refinement. By numerically propagating the reconstructed illumination function, we have obtained the X-ray wavefield profile of the 23 nm round focus of the Fresnel zone plate (outermost zone width, Δr = 20 nm) as well as the X-ray wavefront at the exit pupil of the lens. The measurements of the wavefront aberrations were repeatable to within a root mean square error of 0.006 waves, and we demonstrate that they can be related to manufacturing aspects of the diffractive optical element and to errors on the incident X-ray wavefront introduced by the upstream beamline optics.

Phase tomography from x-ray coherent diffractive imaging projections.

Coherent diffractive imaging provides accurate phase projections that can be tomographically combined to yield detailed quantitative 3D reconstructions with a resolution that is not limited by imaging optics. We present robust algorithms for post-processing and alignment of these tomographic phase projections. A simple method to remove undesired constant and linear phase terms on the reconstructions is given. Also, we provide an algorithm for automatic alignment of projections that has good performance even for samples with no fiducial markers. Currently applied to phase projections, this alignment algorithm has proven to be robust and should also be useful for lens-based tomography techniques that pursue nanoscale 3D imaging. Lastly, we provide a method for tomographic reconstruction that works on phase projections that are known modulo 2π, such that the phase unwrapping step is avoided. We demonstrate the performance of these algorithms by 3D imaging of bacteria population in legume root-nodule cells.

Beam hardening effects in grating-based x-ray phase-contrast imaging.

PURPOSE: In this work, the authors investigate how beam hardening affects the image formation in x-ray phase-contrast imaging and consecutively develop a correction algorithm based on the results of the analysis. METHODS: The authors' approach utilizes a recently developed x-ray imaging technique using a grating interferometer capable of visualizing the differential phase shift of a wave front traversing an object. An analytical description of beam hardening is given, highlighting differences between attenuation and phase-contrast imaging. The authors present exemplary beam hardening artifacts for a number of well-defined samples in measurements at a compact laboratory setup using a polychromatic source. RESULTS: Despite the differences in image formation, the authors show that beam hardening leads to a similar reduction of image quality in phase-contrast imaging as in conventional attenuation-contrast imaging. Additionally, the authors demonstrate that for homogeneous objects, beam hardening artifacts can be corrected by a linearization technique, applicable to all kinds of phase-contrast methods using polychromatic sources. CONCLUSIONS: The evaluated correction algorithm is shown to yield good results for a number of simple test objects and can thus be advocated in medical imaging and nondestructive testing.

Concentration effects on the dynamics of liquid crystalline self-assembly: time-resolved X-ray scattering studies.

A manifold of ordering transitions relevant to chemical and biological systems occur at interfaces from liquids to self-assembled soft solids like membranes or liquid crystals. In the present case, we were interested in understanding the phase transition from the microemulsion phase to the liquid crystal phase in terms of their driving forces, i.e., activation energy and entropy. The purpose of this work was to clarify the influence of concentration effects of the amphiphilic molecules on the nature of these self-assembly processes. By photosensitization of the model system (polyalkylglycolether (C(10)E(4)), water, decane, and cyclohexane) with laser dyes, we could effectively induce and control the phase transition through the absorption of optical photons. The photo transformation conditions were chosen in such a way that the system was in thermal equilibrium. By application of time-resolved photo small-angle X-ray scattering we could monitor the conversion process and demonstrate that the surfactant concentration has a direct impact on the activation energy, which is observable through the length of the induction time.

Nanostructure of healthy and caries-affected human teeth.

Spatially resolved small-angle x-ray scattering based on synchrotron radiation combines the quantitative assessment of nanometer-sized components using scattering with the real-space imaging by means of scanning. The method enables us to study the effect of caries-induced damages on the inorganic and organic nanoscopic components in human teeth. We demonstrate for several 200- to 500-µm-thin tooth slices that the bacterial processes dissolve the ceramic components in enamel and dentin, but the dentinal collagen network remains practically unaffected with respect to its abundance and orientation in early stages of caries and in parts of extended carious lesions. Consequently, we speculate that future caries treatments can be developed reversing the effect of bacterial attacks by means of suitable remineralization of the dentin. FROM THE CLINICAL EDITOR: In this groundbreaking study of caries pathology using synchrotron-based X-ray scattering, the authors demonstrated that while bacterial processes do dissolve the ceramic components in enamel and dentin; however, the dentinal collagen network remains unaffected, enabling the development of future caries treatments that re-mineralize the dentin.

Time scale of glycation in collagen of bovine pericardium-derived bio-tissues.

Glycosyl-ation is the process of combining one or more glucose molecules (or other monosaccharides) with molecules of a different nature (which are therefore glycosyl-ated). In biochemistry, glycosyl-ation is catalyzed by several specific enzymes, and assumes considerable importance since it occurs mainly at the expense of proteins and phospho-lipids which are thus transformed into glycoproteins and glycolipids. Conversely, in diabetes and aging, glycation of proteins is a phenomenon of non-enzymatic nature and thus not easily controlled. Glycation of collagen distorts its structure, renders the extracellular matrix stiff and brittle and at the same time lowers the degradation susceptibility thereby preventing renewal. Based on models detailed in this paper and with parameters determined from experimental data, we describe the glycation of type 1 collagen in bovine pericardium derived bio-tissues, upon incubation in glucose and ribose. With arginine and lysine/hy-droxy-lysine amino acids as the primary sites of glycation and assuming that the topological polar surface area of the sugar molecules determines the glycation rates, we modelled the glycation as a function of time and determined the glycation rate and thus the progression of glycation as well as the resulting volume increase.