Synchrotron Radiation Refraction-Contrast Computed Tomography Based on X-ray Dark-Field Imaging Optics of Pulmonary Malignancy: Comparison with Pathologic Examination.

Refraction-contrast computed tomography based on X-ray dark-field imaging (XDFI) using synchrotron radiation (SR) has shown superior resolution compared to conventional absorption-based methods and is often comparable to pathologic examination under light microscopy. This study aimed to investigate the potential of the XDFI technique for clinical application in lung cancer diagnosis. Two types of lung specimens, primary and secondary malignancies, were investigated using an XDFI optic system at beamline BL14B of the High-Energy Accelerator Research Organization Photon Factory, Tsukuba, Japan. Three-dimensional reconstruction and segmentation were performed on each specimen. Refraction-contrast computed tomographic images were compared with those obtained from pathological examinations. Pulmonary microstructures including arterioles, venules, bronchioles, alveolar sacs, and interalveolar septa were identified in SR images. Malignant lesions could be distinguished from the borders of normal structures. The lepidic pattern was defined as the invasive component of the same primary lung adenocarcinoma. The SR images of secondary lung adenocarcinomas of colorectal origin were distinct from those of primary lung adenocarcinomas. Refraction-contrast images based on XDFI optics of lung tissues correlated well with those of pathological examinations under light microscopy. This imaging method may have the potential for use in lung cancer diagnosis without tissue damage. Considerable equipment modifications are crucial before implementing them from the lab to the hospital in the near future.

Crystal optics simulations for delineation of the three-dimensional cellular nuclear distribution using analyzer-based refraction-contrast computed tomography.

Refraction-contrast computed tomography (RCT) using a refractive angle analyzer of Si perfect crystal can reconstruct the three-dimensional structure of biological soft tissue with contrast comparable to that of stained two-dimensional pathological images. However, the blurring of X-ray beam by the analyzer has prevented improvement of the spatial resolution of RCT, and the currently possible observation of tissue structure at a scale of approximately 20 µm provides only limited medical information. As in pathology, to differentiate between benign and malignant forms of cancer, it is necessary to observe the distribution of the cell nucleus, which is approximately 5-10 µm in diameter. In this study, based on the X-ray dynamical diffraction theory using the Takagi-Taupin equation, which calculates the propagation of X-ray energy in crystals, an analyzer crystal optical system depicting the distribution of cell nuclei was investigated by RCT imaging simulation experiments in terms of the thickness of the Laue-case analyzer, the camera pixel size and the difference in spatial resolution between the Bragg-case and Laue-case analyzers.

Ring artifact removal for differential phase-contrast X-ray computed tomography using a conditional generative adversarial network.

PURPOSE: The integration process used as a pre-processing step in the reconstruction of differential phase-contrast X-ray CT (d-PCCT) causes the measurement noise to propagate throughout the projection image, which is leading to increased ring artifacts (RA) in the reconstructed image. It is difficult to eliminate the RA using conventional RA removal methods that were developed for the absorption-based CT field. We propose an effective method that can remove RA of d-PCCT images. METHODS: The proposed method uses Laplacian images reconstructed from second-derivative projections of d-PCCT. This method is based on a conditional generative adversarial network (cGAN), whose loss function is designed by adding the L1- and L2-norm to the original cGAN. The training data were taken from a numerical phantom generated by a d-PCCT imaging simulator. To validate the applicability of the trained network, we tested its RA removal effect on test data from numerical phantoms generated randomly and actual experimental data. RESULTS: The results of numerical validation using numerical phantoms showed that the proposed method improved the RA removal effect compared to conventional methods. In addition, image comparison by visual evaluation showed that only the proposed method was able to remove RA while preserving original structures in the actual biological d-PCCT images. CONCLUSION: We proposed a cGAN-based method for RA removal that exploits the physical properties of d-PCCT. The proposed method was able to completely remove RA from d-PCCT images on both simulated data and biological data. We believe that this method is useful for the observation of various types of biological soft tissue.

Usefulness of X-ray dark-field imaging in the evaluation of local recurrence after nipple-sparing mastectomy.

PURPOSE: Our previous study suggests that the cross-sectional morphology of ducts and branching of ducts in the nipple are associated with the presence of breast cancer. In this study, we evaluated whether cross-sectional morphology and duct branching of human nipple obtained by X-ray dark-field imaging tomographic technique (XDFI-CT) could predict the likelihood of the presence of intraductal cancer into the nipple. METHODS: A total of 51 nipple specimens were obtained from consecutive total mastectomies performed for breast cancer in Nagoya Medical Center. After reconstructing 3D images of the nipple using XDFI-CT, the cross-sectional images and the 3D arrangement of ducts were extracted. These cross-sectional images of ducts were classified into four patterns based on the status of the lumen without being informed of pathology results. RESULTS: Of the four patterns, the distended ducts with heterogenous content were highly correlated with the presence of ductal carcinoma in situ confirmed by histopathology. The total number of orifices identified in the 51 specimens was 1298, and 182 (14%) at the tip and 19 (1.5%) at least 5 mm depth from the tip were composed of two or more ducts. CONCLUSIONS: Anatomy of nipple ducts is essential to evaluate risk of local recurrence after nipple-sparing mastectomy because cancerous spread occurs within the duct of the same segment of the mammary duct-lobular system in the in situ stage. The 3D microscale anatomy of nipple ducts revealed by XDFI-CT provides useful information to assess the risk of breast cancer involvement at the preserved portion in nipple-sparing mastectomy.

Synchrotron Radiation-Based Refraction-Contrast Tomographic Images Using X-ray Dark-Field Imaging Optics in Human Lung Adenocarcinoma and Histologic Correlations.

The aim of this study was to evaluate the clinical implication of synchrotron radiation imaging techniques for human lung adenocarcinoma in comparison with pathologic examination. A refraction-based tomographic imaging technique called the X-ray dark-field imaging (XDFI) method was used to obtain computed tomographic images of human lung adenocarcinoma at the beam line at Photon Factory BL 14B at the High Energy Accelerator Research Organization (KEK) in Tsukuba, Japan. Images of normal lung tissue were also obtained using the same methods and reconstructed as 3D images. Both reconstructed images were compared with pathologic examinations from histologic slides which were made with identical samples. Pulmonary alveolar structure including terminal bronchioles, alveolar sacs, and vasculatures could be identified in synchrotron radiation images of normal lung. Hyperplasia of interstitial tissue and dysplasia of alveolar structures were noticed in images of lung adenocarcinoma. Both synchrotron radiation images were considerably correlated with images from histologic slides. Lepidic patterns of cancer tissue were distinguished from the invasive area in synchrotron radiation images of lung adenocarcinoma. Refraction-contrast tomographic techniques using synchrotron radiation could provide high-resolution images of lung adenocarcinoma which are compatible with those from pathologic examinations.

X-ray Dark-Field Imaging (XDFI)-a Promising Tool for 3D Virtual Histopathology.

X-ray dark-field imaging (XDFI) utilizing a thin silicon crystal under Laue case enables visualizing three-dimensional (3D) morphological alterations of human tissue. XDFI uses refraction-contrast derived from phase shift rather than absorption as the main X-ray image contrast source to render 2D and 3D images of tissue specimens in unprecedented detail. The unique features of XDFI are its extremely high sensitivity (approximately 1000:1 compared to absorption for soft tissues under X-ray energy of around 20 keV, theoretically) and excellent resolution (8.5 µm) without requiring contrast medium or staining. Thus, XDFI-computed tomography can generate 3D virtual histological images equivalent to those of stained histological sections pathologists observe under low-power light microscopy as far as organs and tissues selected as samples in preliminary studies. This paper reviews the fundamental principles and the potential of XDFI, describes two optical setups for XDFI with examples, illustrates features of XDFI that are salient for histopathology, and presents XDFI examples of refraction-contrast images of atherosclerotic plaques, musculoskeletal tissue, neuronal tissue, and breast cancer specimens. Availability of this X-ray imaging in routine histopathological evaluations of tissue specimens would help guide clinical decision making by highlighting suspicious areas in unstained, thick sections for further sampling and analysis using conventional histopathological techniques. XDFI is a promising tool for 3D virtual histopathology.

Calculation of Stopping-Power Ratio from Multiple CT Numbers Using Photon-Counting CT System: Two- and Three-Parameter-Fitting Method.

The two-parameter-fitting method (PFM) is commonly used to calculate the stopping-power ratio (SPR). This study proposes a new formalism: a three-PFM, which can be used in multiple spectral computed tomography (CT). Using a photon-counting CT system, seven rod-shaped samples of aluminium, graphite, and poly(methyl methacrylate) (PMMA), and four types of biological phantom materials were placed in a water-filled sample holder. The X-ray tube voltage and current were set at 150 kV and 40 µA, respectively, and four CT images were obtained at four threshold settings. A semi-empirical correction method that corrects the difference between the CT values from the photon-counting CT images and theoretical values in each spectral region was also introduced. Both the two- and three-PFMs were used to calculate the effective atomic number and electron density from multiple CT numbers. The mean excitation energy was calculated via parameterisation with the effective atomic number, and the SPR was then calculated from the calculated electron density and mean excitation energy. Then, the SPRs from both methods were compared with the theoretical values. To estimate the noise level of the CT numbers obtained from the photon-counting CT, CT numbers, including noise, were simulated to evaluate the robustness of the aforementioned PFMs. For the aluminium and graphite, the maximum relative errors for the SPRs calculated using the two-PFM and three-PFM were 17.1% and 7.1%, respectively. For the PMMA and biological phantom materials, the maximum relative errors for the SPRs calculated using the two-PFM and three-PFM were 5.5% and 2.0%, respectively. It was concluded that the three-PFM, compared with the two-PFM, can yield SPRs that are closer to the theoretical values and is less affected by noise.

X-ray dark-field phase-contrast imaging: Origins of the concept to practical implementation and applications.

The basic idea of X-ray dark-field imaging (XDFI), first presented in 2000, was based on the concepts used in an X-ray interferometer. In this article, we review 20 years of developments in our theoretical understanding, scientific instrumentation, and experimental demonstration of XDFI and its applications to medical imaging. We first describe the concepts underlying XDFI that are responsible for imparting phase contrast information in projection X-ray images. We then review the algorithms that can convert these projection phase images into three-dimensional tomographic slices. Various implementations of computed tomography reconstructions algorithms for XDFI data are discussed. The next four sections describe and illustrate potential applications of XDFI in pathology, musculoskeletal imaging, oncologic imaging, and neuroimaging. The sample applications that are presented illustrate potential use scenarios for XDFI in histopathology and other clinical applications. Finally, the last section presents future perspectives and potential technical developments that can make XDFI an even more powerful tool.

Improving contrast and spatial resolution in crystal analyzer-based x-ray dark-field imaging: Theoretical considerations and experimental demonstration.

PURPOSE: This paper describes and experimentally validates a methodology for improving contrast and spatial resolution of the x-ray dark-field imaging (XDFI) by cutting the monochromator-collimator asymmetrically and thinning the Laue angle analyzer. METHODS: We measure the spatial resolution of our XDFI setup using a test object consisting of wolfram tungsten meshes and compare it with the theoretical prediction. Using x-ray dark-field computed tomography of breast cancer specimens (lobular carcinoma in situ), we demonstrate that the resolution of XDFI is sufficient for histopathologic analysis. RESULTS: Our experimental results show that the overall spatial resolution of XDFI can be improved by approximately a factor of 2 when these modifications are implemented. The reconstructed images of breast cancer specimens provide sufficient details for radiologic histopathology. CONCLUSIONS: By cutting the monochromator-collimator and thinning the Laue angle analyzer, XDFI can achieve the resolution sufficient for radiologic histopathology.

Cardiac fiber tracking on super high-resolution CT images: a comparative study.

Purpose: High-resolution cardiac imaging and fiber analysis methods are required to understand cardiac anatomy. Although refraction-contrast x-ray CT (RCT) has high soft tissue contrast, it cannot be commonly used because it requires a synchrotron system. Microfocus x-ray CT ( µ CT ) is another commercially available imaging modality. Approach: We evaluate the usefulness of µ CT for analyzing fibers by quantitatively and objectively comparing the results with RCT. To do so, we scanned a rabbit heart by both modalities with our original protocol of prepared materials and compared their image-based analysis results, including fiber orientation estimation and fiber tracking. Results: Fiber orientations estimated by two modalities were closely resembled under the correlation coefficient of 0.63. Tracked fibers from both modalities matched well the anatomical knowledge that fiber orientations are different inside and outside of the left ventricle. However, the µ CT volume caused incorrect tracking around the boundaries caused by stitching scanning. Conclusions: Our experimental results demonstrated that µ CT scanning can be used for cardiac fiber analysis, although further investigation is required in the differences of fiber analysis results on RCT and µ CT .

Three-dimensional microanatomy of human nipple visualized by X-ray dark-field computed tomography.

PURPOSE: The three-dimensional (3D) structure of the human nipple has not been fully clarified. However, its importance has increased in recent years because it has become common practice to preoperatively explore the spread of breast cancer to the nipple with needle biopsy, ductoscopy, and/or ductal lavage for nipple-sparing mastectomy. Here, we demonstrated that X-ray dark-field computed tomography (XDFI-CT) is a powerful tool for reconstructing the 3D distribution pattern of human lactiferous ducts non-destructively, without contrast agent, and with high tissue contrast. METHODS: Nipples amputated from mastectomy specimens of 51 patients with breast cancer were visualized three-dimensionally by XDFI-CT. First, CT images and conventionally stained tissue sections were compared to demonstrate that XDFI-CT provides 3D anatomical information. Next, the number of ducts in the nipple and the number of ducts sharing an ostium near the tip of the nipple were measured from the volume set of XDFI-CT. Finally, the 3D distribution pattern of the ducts was determined. RESULTS: XDFI-CT can provide images almost equivalent to those of low-magnification light microscopy of conventional hematoxylin-eosin-stained histological sections. The mean number of ducts in all cases was 28.0. The total number of ducts sharing an ostium near the tip of the nipple was 525 of 1428. The 3D distribution patterns of the ducts were classified into three types that we defined as convergent (22%), straight (39%), or divergent (39%). CONCLUSIONS: XDFI-CT is useful for exploring the microanatomy of the human nipple and might be used for non-invasive nipple diagnosis in the future.

A carbon CT system: how to obtain accurate stopping power ratio using a Bragg peak reduction technique.

In this study, we investigate the performance of the Gunma University Heavy Ion Medical Center's ion computed tomography (CT) system, which measures the residual range of a carbon-ion beam using a fluoroscopy screen, a charge-coupled-device camera, and a moving wedge absorber and collects CT reconstruction images from each projection angle. Each 2D image was obtained by changing the polymethyl methacrylate (PMMA) thickness, such that all images for one projection could be expressed as the depth distribution in PMMA. The residual range as a function of PMMA depth was related to the range in water through a calibration factor, which was determined by comparing the PMMA-equivalent thickness measured by the ion CT system to the water-equivalent thickness measured by a water column. Aluminium, graphite, PMMA, and five biological phantoms were placed in a sample holder, and the residual range for each was quantified simultaneously. A novel method of CT reconstruction to correct for the angular deflection of incident carbon ions in the heterogeneous region utilising the Bragg peak reduction (BPR) is also introduced in this paper, and its performance is compared with other methods present in the literature such as the decomposition and differential methods. Stopping power ratio values derived with the BPR method from carbon-ion CT images matched closely with the true water-equivalent length values obtained from the validation slab experiment.

Imaging with ultra-small-angle X-ray scattering using a Laue-case analyzer and its application to human breast tumors.

PURPOSE: In this study, we demonstrate a novel imaging technique, based on ultra-small-angle X-ray scattering (USAXS) that uses a Laue-case Si wafer as the angle analyzer. METHODS: We utilized the (1 1 1) diffraction plane of a 356 µm thick, symmetrically cut Si wafer as the angle analyzer, denoted by A[L]. With this device, we performed USAXS imaging experiments using 19.8 keV synchrotron X-rays. The objects we imaged were formalin-fixed, paraffin-embedded breast tumors (an invasive carcinoma and an intraductal papilloma). During image acquisition by a charge-coupled device (CCD) camera, we varied the rotation angle of the analyzer in 0.02″ steps from -2.40″ to +2.40″ around the Bragg angle. The exposure time for each image was 2 s. We determined the amount of ultra-small-angle X-ray scattering from the width of the intensity curve obtained for each local pixel during the rotation of the analyzer. RESULTS: We acquired USAXS images of malignant and benign breast tumor specimens using the A[L] analyzer; regions with larger USAXS form brighter areas in the image. We varied the sensitivity of the USAXS image by changing the threshold level of the object rocking curve. CONCLUSIONS: The USAXS images can provide information about the internal distribution of closely packed scattering bodies in a sample with reasonable sensitivity. This information differs from that obtainable through refraction-contrast imaging. Although further validation studies will be necessary, we conclude that USAXS imaging using a Laue-case analyzer may have significant potential as a new diagnosis technique.

Multi-pinhole fluorescent x-ray computed tomography for molecular imaging.

We propose a multi-pinhole fluorescent x-ray computed tomography (mp-FXCT) technique for preclinical molecular imaging that can provide the complete data necessary to produce 3-D tomographic images during anaesthesia. In this method, multiple projections are simultaneously acquired through a multi-pinhole collimator with a 2-D detector and full-field volumetric beam to accelerate the data acquisition process and enhance the signal-to-noise ratios of the projections. We constructed a 15-pinhole mp-FXCT imaging system at beamline ARNE-7A at KEK and performed preliminary experiments to investigate its imaging properties using physical phantoms and a non-radioactive I imaging agent. The mp-FXCT system could detect an I concentration of 0.038 mg/ml, the minimum required for in-vivo imaging, at a spatial resolution of about 0.3 mm during a data acquisition time of 90 min, which is less than the time for which anaesthesia is effective and suggests that preclinical molecular imaging is feasible with mp-FXCT.

Dual-energy fluorescent x-ray computed tomography system with a pinhole design: Use of K-edge discontinuity for scatter correction.

We propose a pinhole-based fluorescent x-ray computed tomography (p-FXCT) system with a 2-D detector and volumetric beam that can suppress the quality deterioration caused by scatter components. In the corresponding p-FXCT technique, projections are acquired at individual incident energies just above and below the K-edge of the imaged trace element; then, reconstruction is performed based on the two sets of projections using a maximum likelihood expectation maximization algorithm that incorporates the scatter components. We constructed a p-FXCT imaging system and performed a preliminary experiment using a physical phantom and an I imaging agent. The proposed dual-energy p-FXCT improved the contrast-to-noise ratio by a factor of more than 2.5 compared to that attainable using mono-energetic p-FXCT for a 0.3 mg/ml I solution. We also imaged an excised rat's liver infused with a Ba contrast agent to demonstrate the feasibility of imaging a biological sample.

Shear Wave Imaging of Breast Tissue by Color Doppler Shear Wave Elastography.

Shear wave elastography is a distinctive method to access the viscoelastic characteristic of the soft tissue that is difficult to obtain by other imaging modalities. This paper proposes a novel shear wave elastography [color Doppler shear wave imaging (CD SWI)] for breast tissue. Continuous shear wave is produced by a small lightweight actuator, which is attached to the tissue surface. Shear wave wavefront that propagates in tissue is reconstructed as a binary pattern that consists of zero and the maximum flow velocities on color flow image (CFI). Neither any modifications of the ultrasound color flow imaging instrument nor a high frame rate ultrasound imaging instrument is required to obtain the shear wave wavefront map. However, two conditions of shear wave displacement amplitude and shear wave frequency are needed to obtain the map. However, these conditions are not severe restrictions in breast imaging. This is because the minimum displacement amplitude is [Formula: see text] for an ultrasonic wave frequency of 12 MHz and the shear wave frequency is available from several frequencies suited for breast imaging. Fourier analysis along time axis suppresses clutter noise in CFI. A directional filter extracts shear wave, which propagates in the forward direction. Several maps, such as shear wave phase, velocity, and propagation maps, are reconstructed by CD SWI. The accuracy of shear wave velocity measurement is evaluated for homogeneous agar gel phantom by comparing with the acoustic radiation force impulse method. The experimental results for breast tissue are shown for a shear wave frequency of 296.6 Hz.

A Color-Doppler Shear-Wave-Imaging Phase-reconstruction Method Using Four Color Flow Images.

This study investigates shear wave phase map reconstruction using a limited number of color flow images (CFIs) acquired with a color Doppler ultrasound imaging instrument. We propose an efficient reconstruction method to considerably reduce the number of CFIs required for reconstruction and compare this method with Fourier analysis-based color Doppler shear wave imaging. The proposed method uses a two-step phase reconstruction process, including an initial phase map derived from four CFIs using an advanced iterative algorithm of optical interferometry. The second step reduces phase artifacts in the initial phase map using an iterative correction procedure that cycles between the Fourier and inverse Fourier domains while imposing directional filtering and total variation regularization. We demonstrate the efficacy of this method using synthetic and experimental data of a breast phantom and human breast tissue. Our results show that the proposed method maintains image quality and reduces the number of CFIs required to four; previous methods have required at least 32 CFIs to achieve equivalent image quality. The proposed method is applicable to real-time shear wave elastography using a continuous shear wave produced by a mechanical vibrator.

Dark-Field Imaging: Recent developments and potential clinical applications.

This paper describes an X-ray phase contrast imaging technique using analyzer-based optics called X-ray Dark-Field Imaging that has been under development for the past 10years. We describe the theory behind XDFI, the X-ray optics required for implementing it in practice, and algorithms used for 2D, 2.5D, and 3D image reconstruction. The XDFI optical chain consists of an asymmetrically cut, Bragg-type monochromator-collimator that provides a planar monochromatic X-ray beam, a positioning stage for the specimens, a Laue-case angle analyzer, and one or two cameras to capture the dark and bright field images. We demonstrate the soft-tissue discrimination capabilities of XDFI by reconstructing images with absorption and phase contrast. By using a variety of specimens such as breast tissue with cancer, joints with articular cartilage, ex-vivo human eye specimen, and others, we show that refraction-based contrast derived from XDFI is more effective in characterizing anatomical features, articular pathology, and neoplastic disease than conventional absorption-based images. For example, XDFI of breast tissue can discriminate between the normal and diseased terminal duct lobular unit, and between invasive and in-situ cancer. The final section of this paper is devoted to potential future developments to enable clinical and histo-pathological applications of this technique.

In Vitro Validation of an Artefact Suppression Algorithm in X-Ray Phase-Contrast Computed Tomography.

X-ray phase-contrast tomography can significantly increase the contrast-resolution of conventional attenuation-contrast imaging, especially for soft-tissue structures that have very similar attenuation. Just as in attenuation-based tomography, phase contrast tomography requires a linear dependence of aggregate beam direction on the incremental direction alteration caused by individual voxels along the path of the X-ray beam. Dense objects such as calcifications in biological specimens violate this condition. There are extensive beam deflection artefacts in the vicinity of such structures because they result in large distortion of wave front due to the large difference of refractive index; for such large changes in beam direction, the transmittance of the silicon analyzer crystal saturates and is no longer linearly dependent on the angle of refraction. This paper describes a method by which these effects can be overcome and excellent soft-tissue contrast of phase tomography can be preserved in the vicinity of such artefact-producing structures.

Limited view reconstruction for differential phase-contrast computed tomography.

This paper describes an algebraic reconstruction algorithm that uses total variation (TV) regularization for differential phase contrast computed tomography (DPC-CT) using a limited number of views. In order to overcome over-flattening inherent in TV regularization, a two-step reconstruction process is used: we first reconstruct tomographic images of gradient refractive index from differential projections with TV regularization; these images are then used to compute tomographic images of refractive index by solving the Poisson equation. We incorporate TV regularization in the reconstruction process because the distribution of gradient refractive index is much more flattened than the refractive index. Simulations of the proposed method demonstrate that it can achieve satisfactory image quality from a much smaller number of projections than is required by the Nyquist sampling theorem. We experimentally prove the feasibility of the proposed method using dark field imaging optics at PF-14C beamline at the Photon Factory, KEK. The differential phase contrast projection data was experimentally acquired from a biological sample and DPC-CT images were reconstructed. We show that far fewer projections are needed when the proposed algorithm is used.