Nondestructive Three-dimensional Imaging Research Articles

Three-dimensional isotropic imaging of live suspension cells enabled by droplet microvortices

Fast, nondestructive three-dimensional (3D) imaging of live suspension cells remains challenging without substrate treatment or fixation, precluding scalable single-cell morphometry with minimal alterations. While optical sectioning techniques achieve 3D live cell imaging, lateral versus depth resolution differences further complicate analysis. We present a scalable microfluidic method capable of 3D fluorescent isotropic imaging of live, nonadherent cells suspended inside picoliter droplets with high-speed single-cell volumetric readout (800 to 1,200 slices in 5 to 8 s) and near-diffraction limit resolution (~216 nm). The platform features a droplet trap array that leverages flow-induced droplet interfacial shear to generate intradroplet microvortices, which rotate single cells on their axis to enable optical projection tomography (OPT)-based imaging. This allows gentle (~1 mPa shear stress) observation of cells encapsulated inside nontoxic isotonic buffer droplets, facilitating scalable OPT acquisition by simultaneous spinning of hundreds of cells. We demonstrate 3D imaging of live myeloid and lymphoid cells in suspension, including K562 cells, as well as naive and activated T cells-small cells prone to movement in their suspended phenotype. Our fully suspended, orientation-independent cell morphometry, driven by isotropic imaging and spherical harmonic analysis, enabled the study of primary T cells across various immunological activation states. This approach unveiled six distinct nuclear content distributions, contrasting with conventional 2D images that typically portray spheroid and bean-like nuclear shapes associated with lymphocytes. Our arrayed-droplet OPT technology is capable of isotropic, single live-cell 3D imaging, with the potential to perform large-scale morphometry of immune cell effector function states while providing compatibility with microfluidic droplet operations.

Non-destructive three-dimensional imaging of artificially degraded CdS paints by pump-probe microscopy

Cadmium sulfide (CdS) pigments have degraded in several well-known artworks, but the influence of pigment properties and environmental conditions on the degradation process have yet to be fully understood. Traditional non-destructive analysis techniques primarily focus on macroscopic degradation, whereas microscopic information is typically obtained with invasive techniques that require sample removal. Here, we demonstrate the use of pump-probe microscopy to nondestructively visualize the three-dimensional structure and degradation progress of CdS pigments in oil paints. CdS pigments, reproduced following historical synthesis methods, were reproduced as oil paints and artificially aged by exposure to high relative humidity and light. The degradation of CdS to CdSO4·xH2O was confirmed by both FTIR (Fourier-transform infrared) and XPS (x-ray photoelectron spectroscopy) experiments. During the degradation process, optical pump-probe microscopy was applied to track the degradation progress in single grains, and volumetric imaging revealed early CdS degradation of small particles and on the surface of large particles. This indicates that the particle dimension influences the extent and evolution of degradation of historical CdS. In addition, the pump-probe signal decrease in degraded CdS is observable before visible changes to the eye, demonstrating that pump-probe microscopy is a promising tool to detect early-stage degradation in artworks.

Continuous-wave terahertz in-line holographic diffraction tomography with the scattering fields reconstructed by a physics-enhanced deep neural network

Diffraction tomography is a promising, quantitative, and nondestructive three-dimensional (3D) imaging method that enables us to obtain the complex refractive index distribution of a sample. The acquisition of the scattered fields under the different illumination angles is a key issue, where the complex scattered fields need to be retrieved. Presently, in order to develop terahertz (THz) diffraction tomography, the advanced acquisition of the scattered fields is desired. In this paper, a THz in-line digital holographic diffraction tomography (THz-IDHDT) is proposed with an extremely compact optical configuration and implemented for the first time, to the best of our knowledge. A learning-based phase retrieval algorithm by combining the physical model and the convolution neural networks, named the physics-enhanced deep neural network (PhysenNet), is applied to reconstruct the THz in-line digital hologram, and obtain the complex amplitude distribution of the sample with high fidelity. The advantages of the PhysenNet are that there is no need for pretraining by using a large set of labeled data, and it can also work for thick samples. Experimentally with a continuous-wave THz laser, the PhysenNet is first demonstrated by using the thin samples and exhibits superiority in terms of imaging quality. More importantly, with regard to the thick samples, PhysenNet still works well, and can offer 2D complex scattered fields for diffraction tomography. Furthermore, the 3D refractive index maps of two types of foam sphere samples are successfully reconstructed by the proposed method. For a single foam sphere, the relative error of the average refractive index value is only 0.17%, compared to the commercial THz time-domain spectroscopy system. This demonstrates the feasibility and high accuracy of the THz-IDHDT, and the idea can be applied to other wavebands as well.

Trabecular Bone Microarchitecture Evaluation in an Osteoporosis Mouse Model.

Bone microstructure refers to the arrangement and quality of bone tissue at the microscopic level. Understanding the bone microstructure of the skeleton is crucial for gaining insight into the pathophysiology of osteoporosis and improving its treatment. However, handling bone samples can be complex due to their hard and dense properties. Secondly, specialized software makes image processing and analysis difficult. In this protocol, we present a cost-effective and easy-to-use solution for trabecular bone microstructure analysis. Detailed steps and precautions are provided. Micro-CT is a non-destructive three-dimensional (3D) imaging technique that provides high-resolution images of trabecular bone structure. It allows for the objective and quantitative evaluation of bone quality, which is why it is widely regarded as the gold standard method for bone quality assessment. However, histomorphometry remains indispensable as it offers crucial cellular-level parameters, bridging the gap between two-dimensional (2D) and 3D assessments of bone specimens. As for the histologic techniques, we chose to decalcify the bone tissue and then perform traditional paraffin embedding. In summary, combining these two methods can provide more comprehensive and accurate information on bone microstructure.

Automated detection and growth tracking of 3D bio-printed organoid clusters using optical coherence tomography with deep convolutional neural networks.

Organoids are advancing the development of accurate prediction of drug efficacy and toxicity in vitro. These advancements are attributed to the ability of organoids to recapitulate key structural and functional features of organs and parent tumor. Specifically, organoids are self-organized assembly with a multi-scale structure of 30-800μm, which exacerbates the difficulty of non-destructive three-dimensional (3D) imaging, tracking and classification analysis for organoid clusters by traditional microscopy techniques. Here, we devise a 3D imaging, segmentation and analysis method based on Optical coherence tomography (OCT) technology and deep convolutional neural networks (CNNs) for printed organoid clusters (Organoid Printing and optical coherence tomography-based analysis, OPO). The results demonstrate that the organoid scale influences the segmentation effect of the neural network. The multi-scale information-guided optimized EGO-Net we designed achieves the best results, especially showing better recognition workout for the biologically significant organoid with diameter ≥50μm than other neural networks. Moreover, OPO achieves to reconstruct the multiscale structure of organoid clusters within printed microbeads and calibrate the printing errors by segmenting the printed microbeads edges. Overall, the classification, tracking and quantitative analysis based on image reveal that the growth process of organoid undergoes morphological changes such as volume growth, cavity creation and fusion, and quantitative calculation of the volume demonstrates that the growth rate of organoid is associated with the initial scale. The new method we proposed enable the study of growth, structural evolution and heterogeneity for the organoid cluster, which is valuable for drug screening and tumor drug sensitivity detection based on organoids.

Standardised quantitative ultrasound imaging approach for the contact-less three-dimensional analysis of neocartilage formation in hydrogel-based bioscaffolds

In this work we present a standardised quantitative ultrasound imaging (SQUI) approach for the non-destructive three-dimensional imaging and quantification of cartilage formation in hydrogel based bioscaffolds. The standardised concept involves the processing of ultrasound backscatter data with respect to an acellular phantom in combination with the compensation of sound speed mismatch diffraction effects between the bioscaffold and the phantom. As a proof-of-concept, the SQUI approach was tested on a variety of bioscaffolds with varying degree of neocartilage formation. These were composed of Gelatine Methacryloyl (GelMA) hydrogels laden with human adipose-derived stem cells (hADSCs). These were cultured under chondrogenic stimulation following a previously established protocol, where the degree of the neocartilage formation was modulated using different GelMA network densities (6, 8, 10 % w/v) and culture time (0, 14, 28 days). Using the SQUI approach we were able to detect marked acoustic and morphological changes occurring in the bioscaffolds a result of their different chondrogenic outcome. We defined an acoustic neocartilage indicator, the sonomarker, for the selective imaging and quantification of neocartilage formation. The sonomarker, of backscatter intensity logIBC -2.4, was found to correlate with data obtained via standard destructive bioassays. The ultrasonic evaluation of human specimens confirmed the sonomarker as a relevant intensity, although it was found to shift to higher intensity values in proportion to the cartilage condition as inferred from sound speed measurements. This study demonstrates the potential of the SQUI approach for the realization of non-destructive analysis of cartilage regeneration over-time. STATEMENT OF SIGNIFICANCE: As tissue engineering strategies for neocartilage regeneration evolve towards clinical implementation, alternative characterisation approaches that allow the non-destructive monitoring of extracellular matrix formation in implantable hydrogel based bioscaffolds are needed. In this work we present an innovative standardized quantitative ultrasound imaging (SQUI) approach that allows the non-destructive, volumetric, and quantitative evaluation of neocartilage formation in hydrogel based bioscaffolds. The standardised concept aims to provide a robust approach that accounts for the dynamic changes occurring during the conversion from a cellular bioscaffold towards the formation of a neocartilage construct. We believe that the SQUI approach will be of great benefit for the evaluation of constructs developing neocartilage, not only for in-vitro applications but also potentially applicable to in-vivo applications.

Three-dimensional visualization of nanoparticle lattices and multimaterial frameworks.

Advances in nanoscale self-assembly have enabled the formation of complex nanoscale architectures. However, the development of self-assembly strategies toward bottom-up nanofabrication is impeded by challenges in revealing these structures volumetrically at the single-component level and with elemental sensitivity. Leveraging advances in nano-focused hard x-rays, DNA-programmable nanoparticle assembly, and nanoscale inorganic templating, we demonstrate nondestructive three-dimensional imaging of complexly organized nanoparticles and multimaterial frameworks. In a three-dimensional lattice with a size of 2 micrometers, we determined the positions of about 10,000 individual nanoparticles with 7-nanometer resolution, and identified arrangements of assembly motifs and a resulting multimaterial framework with elemental sensitivity. The real-space reconstruction permits direct three-dimensional imaging of lattices, which reveals their imperfections and interfaces and also clarifies the relationship between lattices and assembly motifs.

Immunofluorescence-guided segmentation of three-dimensional features in micro-computed tomography datasets of human lung tissue.

Micro-computed tomography (µCT) provides non-destructive three-dimensional (3D) imaging of soft tissue microstructures. Specific features in µCT images can be identified using correlated two-dimensional (2D) histology images allowing manual segmentation. However, this is very time-consuming and requires specialist knowledge of the tissue and imaging modalities involved. Using a custom-designed µCT system optimized for imaging unstained formalin-fixed paraffin-embedded soft tissues, we imaged human lung tissue at isotropic voxel sizes less than 10 µm. Tissue sections were stained with haematoxylin and eosin or cytokeratin 18 in columnar airway epithelial cells using immunofluorescence (IF), as an exemplar of this workflow. Novel utilization of tissue autofluorescence allowed automatic alignment of 2D microscopy images to the 3D µCT data using scripted co-registration and automated image warping algorithms. Warped IF images, which were accurately aligned with the µCT datasets, allowed 3D segmentation of immunoreactive tissue microstructures in the human lung. Blood vessels were segmented semi-automatically using the co-registered µCT datasets. Correlating 2D IF and 3D µCT data enables accurate identification, localization and segmentation of features in fixed soft lung tissue. Our novel correlative imaging workflow provides faster and more automated 3D segmentation of µCT datasets. This is applicable to the huge range of formalin-fixed paraffin-embedded tissues held in biobanks and archives.

PEG-modified gadolinium nanoparticles as contrast agents for in vivo micro-CT

Vascular research is largely performed in rodents with the goal of developing treatments for human disease. Micro-computed tomography (micro-CT) provides non-destructive three-dimensional imaging that can be used to study the vasculature of rodents. However, to distinguish vasculature from other soft tissues, long-circulating contrast agents are required. In this study, we demonstrated that poly(ethylene glycol) (PEG)-coated gadolinium nanoparticles can be used as a vascular contrast agent in micro-CT. The coated particles could be lyophilized and then redispersed in an aqueous solution to achieve 100 mg/mL of gadolinium. After an intravenous injection of the contrast agent into mice, micro-CT scans showed blood pool contrast enhancements of at least 200 HU for 30 min. Imaging and quantitative analysis of gadolinium in tissues showed the presence of contrast agent in clearance organs including the liver and spleen and very low amounts in other organs. In vitro cell culture experiments, subcutaneous injections, and analysis of mouse body weight suggested that the agents exhibited low toxicity. Histological analysis of tissues 5 days after injection of the contrast agent showed cytotoxicity in the spleen, but no abnormalities were observed in the liver, lungs, kidneys, and bladder.

Double-exposure method for speckle-tracking x-ray phase-contrast microtomography

X-ray phase-contrast microtomography based on speckle tracking is an attractive method for non-destructive three-dimensional imaging owing to its simple setup and ability to yield absorption, refractive, and scattering images simultaneously. However, the edge-enhancement effect usually results in image artifacts or inaccurate phase retrieval, limiting the extensive application of this method in biomedical research and for low-Z materials. In this paper, a double-exposure method is introduced to solve this problem efficiently and accurately. Pure phase samples with various microstructures and densities and a biological sample with a distinct edge-enhancement effect were used to verify the effectiveness of the developed method. In an experiment performed using a polymer phantom with an evenly distributed density, 17 irregularly shaped particles with diameters ranging from 15 to 25 μm were successfully reconstructed with the effective elimination of the edge-enhancement effect. The results obtained for a sample composed of different polymer materials demonstrated that, in contrast with the traditional speckle-tracking method, the present method is able to discriminate materials with similar x-ray attenuations. Finally, experiments were performed using a dehydrated fish, which entail typical edge enhancement and a complex microstructure; notably, it was verified that the fine structure of the fish, including its fins and intestines, was reconstructed completely using the proposed method, whereas the standard speckle-tracking method was ineffective. In conclusion, the developed double-exposure method can serve as an efficient and accurate technique for the x-ray phase-contrast microtomography of samples comprising low-Z materials and complicated microstructures.

Acoustic subsurface-atomic force microscopy: Three-dimensional imaging at the nanoscale

The development of acoustic subsurface atomic force microscopy, which promises three-dimensional imaging with single-digit nanometer resolution by the introduction of ultrasound actuations to a conventional atomic force microscope, has come a long way since its inception in the early 1990s. Recent advances provide a quantitative understanding of the different experimentally observed contrast mechanisms, which paves the way for future applications. In this Perspective, we first review the different subsurface atomic force microscope modalities: ultrasonic force microscopy, atomic force acoustic microscopy, heterodyne force microscopy, mode-synthesizing atomic force microscopy, and near-field picosecond ultrasonic microscopy. Then, we highlight and resolve a debate existing in the literature on the importance of the chosen ultrasound excitation frequencies with respect to the resonance frequencies of the cantilever and the observed contrast mechanisms. Finally, we discuss remaining open problems in the field and motivate the importance of new actuators, near-field picosecond ultrasonics, and integration with other techniques to achieve multi-functional non-destructive three-dimensional imaging at the nanoscale.

Testing the X-ray computed microtomography on microfossil identification: An example from Sergipe-Alagoas Basin, Brazil

Micropaleontology is crucial branch of Earth Sciences, with a pivotal role in the success of the oil and gas industry over the decades. This sector of paleontology is based on the taxonomical description of microfossils, which encompass fossils with size variation from 0.001 mm to 1 m. Normally these microorganisms have a high rate on preservation, thus they are widely accepted as reliable evidence to infer about paleodepositional settings and paleoclimate change. Furthermore, microfossils are critical to understanding the evolution through time, being many of them recognized as index fossil and providing useful biozones to correlation. Since 1950, microfossils have been taxonomically described mainly by stereomicroscopes. However, as the technology progress, traditional ways to study these organisms are challenged and improved by nondestructive three-dimensional imaging techniques, as X-ray tomographic microscopy (SRXTM), three-dimensional X-ray microscopy (3D-XRM), the X-ray computed microtomography (microCT) and X-ray computed nanotomography (nanoCT). Recently, one of the most compelling research areas in micropaleontology is the desire to automatize and enhance the details of systematic classification. Thereby, an increasing number of researches have applied the high-resolution X-ray analysis aiming to improve the morphological, taxonomic and taphonomic examination. Most of them have detailed the enhancement provided by the technique when compared with the standard microscopes, and raised questions about the traditional characters used on the microfossil systematic taxonomy. Nevertheless, even with the surpassing detail on microfossil characterization, the application of microCT has been hampered by the costs and sometimes by the needs of specific computer skill. Thus, this research has evaluated the use of microCT as the technique to classify a random bulk of microfossil (comprising foraminifers, ostracods , radiolarians , gastropods and echinoderms) with no further software treatment. No standard microscope analysis was performed. Despite chemical composition of microfossils, most of the specimens morphology, especially internal structures, have been easily acquired and analyzed. 96% of the microfossils of the dataset were identified at least on genus level. Irrespective of the deepness of detail, when considering the overall taxonomic identification, the microCT seems to be effective as the standard microscope. Nonetheless, when problematic specimens are evaluated, the microCT seems to be a more reliable and practical tool than other methods as Scanning Electron Microscopy. Thus, the technique can be used solo or as a complementary method to the stereomicroscope. Additionally, the high-resolution has the potential to lead to the expected automatized micropaleontology, since they can provide numerous images in several planes. This may create a strong database necessary to machine learning and computer identification. • Random bulk of microfossils are evaluated through microCT. • No further computer treatment on microfossil image. • microCT effective method to microfossil identification.

Research on Mechanical Deviation Correction Method for Industrial CT

Industrial CT is the abbreviation of industrial computed tomography (CT) technology, which adopts X-ray imaging principle to realize the non-destructive three-dimensional imaging to obtain high-precision three-dimensional tomographic data and volume data inside the product, clear, accurate and intuitive. In addition, it can show the internal structure, composition, material and defective condition of the inspected object. Factors such as mechanical accuracy and mounting deviations can adversely affect CT imaging and, in severe cases, lead to CT imaging failure. It is important to achieve effective correction based on the existing CT mechanical accuracy and mounting deviations. In this paper the authors collect data and performed experiments in a real CT system, they use standard test blocks to correct the deviations while keeping the existing CT system unchanged, the effectiveness can be validated by the experimental data.

Improvement of the depth resolution of swept-source THz-OCT for non-destructive inspection.

We construct a terahertz swept source optical coherence tomography system using a continuous-wave diode multiplier source in the 600-GHz band for defect inspection in multilayer objects and evaluate its performance. Using this system, we image a multilayer plastic sample to demonstrate the effectiveness of nondestructive three-dimensional imaging. To enhance the depth resolution, we apply an annihilating filter to the analysis and confirm that two surfaces of a 1-mm-thick plastic plate can be resolved. In addition, the repeatability of measured thicknesses is 0.22 mm. These values are approximately one-half and one-tenth of the resolution achievable by conventional Fourier analysis, respectively.

Equally sloped tomography based X-ray full-field nano-CT at Shanghai Synchrotron Radiation Facility

Abstract X-ray full-field nano-computed tomography (nano-CT) has non-destructive three-dimensional imaging capabilities with high spatial resolution, and has been widely applied to investigate morphology and structures in various areas. Conventional tomography reconstructs a 3D object from a large number of equal-angle projections. For nano-CT, it takes long collecting time due to the large projection numbers and long exposure time. Here, equally-sloped tomography (EST) based nano-CT was implemented and constructed on X-ray imaging beamline at the Shanghai Synchrotron Radiation Facility (SSRF) to overcome or alleviate these difficulties. Preliminary results show that hard TXM with the spatial resolution of 100 nm and the EST-based nano-CT with the ability of 3D nano non-destructive characterization have been realized. This technique promotes hard X-ray imaging capability to nano scales at SSRF and could have applications in many fields including nanomaterials, new energy and life sciences. The study will be helpful for the construction of the new full field X-ray nano-imaging beamline with the spatial resolution of 20 nm at SSRF phase II project.

Investigation of an Egyptian Mummy board by Using Clinical Multi-slice Computed Tomography

ABSTRACTA modern approach to the diagnosis of artistic heritage makes use of non-destructive techniques which avoid damage to artworks or signs of interaction with the analytical technique used. For this reason, today, the employment of X-rays to study artistic heritage is quite widespread. Computed tomography (CT) Imaging was first used as specialized medical diagnostic tool in the early 1970s. Today, CT scanners have become more common, and non-destructive three-dimensional imaging has found an increasing number of uses in different areas such as materials research, geology, archaeology, and museum conservation. In this work, a new generation of clinical CT (based on the Dual Energy and multi-slice acquisition principle) is used to investigate an Egyptian wooden mummy board with a total acquisition time of 93.23 seconds and a voxel size of 0.078 × 0.078 × 1.5 mm. The use of two distinct techniques of image post-processing, the two-dimensional multi-planar reconstruction (MPR) and the three-dimensional volume rendering technique (VRT) allowed us to: evaluate the state of conservation of the mummy board, determine how the wooden object was implemented, recognize a nineteenth-century intervention of consolidation, and advance the hypothesis that the wooden planks were reused. In particular, we highlight the use of the cardiovascular and pulmonary filter with VRT reconstruction, which allows a virtual elimination of different parts of the mummy board to investigate those areas otherwise accessible only by removing parts of the board and thus damaging the work.

(Invited) Non-Destructive Three-Dimensional Imaging of Extended Defects in 4H-SiC

Grazing-incidence X-ray topography and photoluminescence (PL) imaging are utilized to understand the nature and behavior of extended defects in 4H-SiC. Although these methods are useful for non-destructive defect analysis, they provide two-dimensional (2D) images with low depth resolution. Advanced three-dimensional (3D) imaging methods are thus expected to be used for characterizing extended defects. In particular, very high-voltage SiC bipolar devices with a thick epilayer require defect imaging as deep as 100-200 μm. We established a 3D imaging technique using an X-ray microbeam and a novel fine slit, which was successfully applied to characterizing threading screw dislocations (TSDs) [1, 2], threading edge dislocations (TEDs), and basal-plane dislocations (BPDs) [3, 4] in 4H-SiC. However, this method requires a large-scale synchrotron-radiation facility. In this study, we show how second-harmonic generation (SHG) and two-photon-excited photoluminescence (2PPL) imaging techniques are powerful tools for 3D analysis of extended defects in 4H-SiC epilayers [5, 6]. The SHG and 2PPL methods can be performed by a multi-photon microscope in individual laboratories. The SHG method provides clear 3D images of 3C-inclusions because 3C-SiC is SHG-active, but not 4H-SiC host crystal in c-axis incidence (Fig. 1). The 2PPL method yields 3D images; not only of 3C-inclusions but also 8H stacking faults (Fig. 2), and single Shockley stacking faults in the epilayers. The 2PPL method also achieved 3D imaging of TSDs, TEDs, and BPDs using band-edge emission [7]. Since band-edge emission quenches near defects, these dislocations can be visualized as dark contrasts on a bright background. We obtained 3D images of TSDs and TEDs extending ~200 μm from the surface (Fig. 3). Unlike 2D PL imaging with uniform illumination, the dark-contrast imaging is governed by the diffusion and transport of excess carriers injected from a scanning focal point. A simulation study was conducted to reveal the mechanism of the dark-contrast imaging. Each of the dislocations provided 2D dark-contrast images stacked depthwise, from which the tilt angles of dislocations were determined. Figure 4 shows the plots of φ versus θ for 89 TSDs in the 4º off epilayer, where θ denotes the angle of the dislocation line from the c-axis, and φ the counter-clockwise angle of (0 0 0 1) projected dislocation lines from the step-flow [1 1 -2 0] direction. Their dislocation lines incline in the step-flow direction. It is also shown that the regions of left- and right-handed (LH and RH) dislocations clearly differ. The plots in Fig. 4 include those of 1c and c+a dislocations. We found that 1c dislocations had very similar tilt angles, whereas those of c+a dislocations were largely spread out. The tilt angles were also investigated for 105 TSDs in the 8º off epilayer, which exhibits larger θ angles and a smaller φ range than the 4º off-cut case. We examined the tilt angles of more than 300 TEDs and found that the TEDs not only inclined in the step-flow direction, but also the direction of the extra half planes [7]. The results of the tilt-angle analyses cannot be explained by the energy minimization model in bulk materials. We consider that the directions of TEDs and TSDs depend upon the interactions between dislocations and advancing steps on a growing surface. [1] R. Tanuma, T. Kubo, F. Togoh, T. Tawara, A. Saito, K. Fukuda, K. Hayashi, and Y. Tsusaka, Phys. Status Solidi A 204, 2706 (2007). [2] R. Tanuma, T. Tamori, Y. Yonezawa, H. Yamaguchi, H. Matsuhata, K. Fukuda, and K. Arai, Material Sci. Forum 615-617, 251 (2009). [3] R. Tanuma, D. Mori, I. Kamata, and H. Tsuchida, Appl. Phys. Express 5, 061301 (2012). [4] R. Tanuma, D. Mori, I. Kamata, and H. Tsuchida, J. Appl. Phys. 114, 023511 (2013). [5] R. Tanuma and H. Tsuchida, Appl. Phys. Express 7, 021304 (2014). [6] R. Tanuma and H. Tsuchida, Mat. Sci. Forum 778-780, 338 (2014). [7] R. Tanuma, M. Nagano, I. Kamata, and H. Tsuchida, Appl. Phys. Express 7, 121303 (2014). Figure 1

(Invited) Non-Destructive Three-Dimensional Imaging of Extended Defects in 4H-SiC

Three-dimensional (3D) imaging of extended defects in 4H-SiC epilayers was achieved by optical second-harmonic generation (SHG) and two-photon-excited photoluminescence (2PPL). The SHG method provided 3D images of 3C-inclusions with a spatial resolution of about 1.3 μm. Using the 2PPL method, we conducted the 3D imaging of 3C-inclusions, 8H stacking faults, and single Shockley stacking faults in the epilayers. The band-edge emission of 2PPL enabled the 3D imaging of threading screw dislocations (TSDs), threading edge dislocations (TEDs), and basal-plane dislocations (BPDs). Since band-edge emission quenches near defects, these dislocations can be visualized as dark contrasts on a bright background. The perspective view of TSDs and TEDs extending ~200 μm from the surface was given by inverting the intensity of a 3D image block of band-edge emission. The tilt angles of TSDs and TEDs in 4H-SiC epilayers were also measured and the mechanisms governing the line directions of TEDs and TSDs were discussed.

Structural and compositional analysis of a casting mold sherd from ancient China.

Casting had symbolic significance and was strictly controlled in the Shang dynasty of ancient China. Vessel casting was mainly distributed around the Shang capital, Yin Ruins, which indicates a rigorous centralization of authority. Thus, for a casting mold to be excavated far from the capital region is rare. In addition to some bronze vessel molds excavated at the Buyao Village site, another key discovery of a bronze vessel mold occurred at Daxinzhuang. The Daxinzhuang site was a core area in the east of Shang state and is an important site to study the eastward expansion of the Shang. Here, combining synchrotron X-rays and other physicochemical analysis methods, nondestructive three-dimensional structure imaging and different elemental analyses were conducted on this mold sherd. Through high penetration X-ray tomography, we obtained insights on the internal structure and discovered some pores. We infer that the generation of pores inside the casting mold sherd was used to enhance air permeability during casting. Furthermore, we suppose that the decorative patterns on the surface were carved and not pasted onto it. Considering the previous compositional studies of bronze vessels, the copper and iron elements were analyzed by different methods. Unexpectedly, a larger amount of iron than of copper was detected on the surface. According to the data analysis and archaeological context, the source of iron on the casting mold sherd could be attributed to local soil contamination. A refined compositional analysis confirms that this casting mold was fabricated locally and used for bronze casting.

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.