Phase contrast micro-CT with adjustable in-slice spatial resolution at constant magnification.

Objective.To report on a micro computed tomography (micro-CT) system capable of x-ray phase contrast imaging and of increasing spatial resolution at constant magnification.Approach.The micro-CT system implements the edge illumination (EI) method, which relies on two absorbing masks with periodically spaced transmitting apertures in the beam path; these split the beam into an array of beamlets and provide sensitivity to the beamlets' directionality, i.e. refraction. In EI, spatial resolution depends on the width of the beamlets rather than on the source/detector point spread function (PSF), meaning that resolution can be increased by decreasing the mask apertures, without changing the source/detector PSF or the magnification.Main results.We have designed a dedicated mask featuring multiple bands with differently sized apertures and used this to demonstrate that resolution is a tuneable parameter in our system, by showing that increasingly small apertures deliver increasingly detailed images. Phase contrast images of a bar pattern-based resolution phantom and a biological sample (a mouse embryo) were obtained at multiple resolutions.Significance.The new micro-CT system could find application in areas where phase contrast is already known to provide superior image quality, while the added tuneable resolution functionality could enable more sophisticated analyses in these applications, e.g. by scanning samples at multiple scales.

Edge-illumination spectral phase-contrast tomography.

Following the rapid, but independent, diffusion of x-ray spectral and phase-contrast systems, this work demonstrates the first combination of spectral and phase-contrast computed tomography (CT) obtained by using the edge-illumination technique and a CdTe small-pixel (62µm) spectral detector. A theoretical model is introduced, starting from a standard attenuation-based spectral decomposition and leading to spectral phase-contrast material decomposition. Each step of the model is followed by quantification of accuracy and sensitivity on experimental data of a test phantom containing different solutions with known concentrations. An example of a micro CT application (20µm voxel size) on an iodine-perfusedex vivomurine model is reported. The work demonstrates that spectral-phase contrast combines the advantages of spectral imaging, i.e. high-Zmaterial discrimination capability, and phase-contrast imaging, i.e. soft tissue sensitivity, yielding simultaneously mass density maps of water, calcium, and iodine with an accuracy of 1.1%, 3.5%, and 1.9% (root mean square errors), respectively. Results also show a 9-fold increase in the signal-to-noise ratio of the water channel when compared to standard spectral decomposition. The application to the murine model revealed the potential of the technique in the simultaneous 3D visualization of soft tissue, bone, and vasculature. While being implemented by using a broad spectrum (pink beam) at a synchrotron radiation facility (Elettra, Trieste, Italy), the proposed experimental setup can be readily translated to compact laboratory systems including conventional x-ray tubes.

X-ray phase-contrast microtomography of soft tissues using a compact laboratory system with two-directional sensitivity.

X-ray microtomography is a nondestructive, three-dimensional inspection technique applied across a vast range of fields and disciplines, ranging from research to industrial, encompassing engineering, biology, and medical research. Phase-contrast imaging extends the domain of application of x-ray microtomography to classes of samples that exhibit weak attenuation, thus appearing with poor contrast in standard x-ray imaging. Notable examples are low-atomic-number materials, like carbon-fiber composites, soft matter, and biological soft tissues. We report on a compact and cost-effective system for x-ray phase-contrast microtomography. The system features high sensitivity to phase gradients and high resolution, requires a low-power sealed x-ray tube, a single optical element, and fits in a small footprint. It is compatible with standard x-ray detector technologies: in our experiments, we have observed that single-photon counting offered higher angular sensitivity, whereas flat panels provided a larger field of view. The system is benchmarked against known-material phantoms, and its potential for soft-tissue three-dimensional imaging is demonstrated on small-animal organs: a piglet esophagus and a rat heart. We believe that the simplicity of the setup we are proposing, combined with its robustness and sensitivity, will facilitate accessing quantitative x-ray phase-contrast microtomography as a research tool across disciplines, including tissue engineering, materials science, and nondestructive testing in general.

Technical note: Cartilage imaging with sub-cellular resolution using a laboratory-based phase-contrast x-ray microscope.

BACKGROUND: Microscopic imaging of cartilage is a key tool for the study and development of treatments for osteoarthritis. When cellular and sub-cellular resolution is required, histology remains the gold standard approach, albeit limited by the lack of volumetric information as well as by processing artifacts. Cartilage imaging with the sub-cellular resolution has only been demonstrated in the synchrotron environment. PURPOSE: To provide a proof-of-concept demonstration of the capability of a laboratory-based x-ray phase-contrast microscope to resolve sub-cellular features in a cartilage sample. METHODS: This work is based on a laboratory-based x-ray microscope using intensity-modulation masks. The structured nature of the beam, resulting from the mask apertures, allows the retrieval of three contrast channels, namely, transmission, refraction and dark-field, with resolution depending only on the mask aperture width. An ex vivo equine cartilage sample was imaged with the x-ray microscope and results were validated with synchrotron tomography and histology. RESULTS: Individual chondrocytes, that is, cells responsible for cartilage formation, could be detected with the laboratory-based microscope. The complementarity of the three retrieved contrast channels allowed the detection of sub-cellular features in the chondrocytes. CONCLUSIONS: We provide the first proof-of-concept of imaging cartilage tissue with sub-cellular resolution using a laboratory-based x-ray microscope.

Modulation transfer function (MTF) evaluation for x-ray phase imaging system employing attenuation masks.

Objective.Attenuation masks can be used in x-ray imaging systems to increase their inherent spatial resolution and/or make them sensitive to phase effects, a typical example being Edge Illumination x-ray phase contrast imaging (EI-XPCI). This work investigates the performance of a mask-based system such as EI-XPCI in terms of Modulation Transfer Function (MTF), in the absence of phase effects.Approach. Pre-sampled MTF measurements, using an edge, were performed on the same system implemented without masks, with non-skipped masks and finally with skipped masks (i.e. masks in which apertures illuminate every other pixel row/column). Results are compared to simulations and finally images of a resolution bar pattern acquired with all the above setups are presented.Main results. Compared to the detector's inherent MTF, the non-skipped mask setup provides improved MTF results. In comparison to an ideal case where signal spill-out into neighbouring pixels is negligible, this improvement takes place only at specific frequencies of the MTF, dictated by the spatial repetition of the spill-out signal. This is limited with skipped masks, which indeed provide further MTF improvements over a larger frequency range. Experimental MTF measurements are supported through simulation and resolution bar pattern images.Significance. This work has quantified the improvement in MTF due to the use of attenuation masks and lays the foundation for how acceptance and routine quality control tests will have to be modified when systems using masks are introduced in clinical practice and how MTF results will compare to those of conventional imaging systems.

Femtosecond multimodal imaging with a laser-driven X-ray source.

Laser-plasma accelerators are compact linear accelerators based on the interaction of high-power lasers with plasma to form accelerating structures up to 1000 times smaller than standard radiofrequency cavities, and they come with an embedded X-ray source, namely betatron source, with unique properties: small source size and femtosecond pulse duration. A still unexplored possibility to exploit the betatron source comes from combining it with imaging methods able to encode multiple information like transmission and phase into a single-shot acquisition approach. In this work, we combine edge illumination-beam tracking (EI-BT) with a betatron X-ray source and present the demonstration of multimodal imaging (transmission, refraction, and scattering) with a compact light source down to the femtosecond timescale. The advantage of EI-BT is that it allows multimodal X-ray imaging technique, granting access to transmission, refraction and scattering signals from standard low-coherence laboratory X-ray sources in a single shot.

Freestanding high-aspect-ratio gold masks for low-energy, phase-based x-ray microscopy.

High-resolution, x-ray phase contrast microscopy, a key technique with promising potential in biomedical imaging and diagnostics, is based on narrow-slit high-aspect-ratio gold gratings. We present the development, fabrication details, and experimental testing of the freestanding 10µm thick gold membrane masks with an array of 0.9-1.5µm void slit apertures for a novel low-energy x-ray microscope. The overall mask size is 4 mm × 4 mm, with a grating pitch of 7.5µm, 6.0-6.6µm wide gold bars are supported by 3µm wide crosslinks at 400µm intervals. The fabrication process is based on gold electroplating into a silicon mold coated with various thin films to form a voltage barrier, plating base, and sacrificial layer, followed by the mold removal to obtain the freestanding gold membrane with void slit apertures. We discuss key aspects for the materials and processes, including gold structures homogeneity, residual stresses, and prevention of collapsing of the grid elements. We further demonstrate the possibility to obtain high-resolution, high contrast 2D images of biological samples using an incoherent, rotating anode x-ray tube.

Time resolved in-situ multi-contrast X-ray imaging of melting in metals.

In this work, the application of a time resolved multi-contrast beam tracking technique to the investigation of the melting and solidification process in metals is presented. The use of such a technique allows retrieval of three contrast channels, transmission, refraction and dark-field, with millisecond time resolution. We investigated different melting conditions to characterize, at a proof-of-concept level, the features visible in each of the contrast channels. We found that the phase contrast channel provides a superior visibility of the density variations, allowing the liquid metal pool to be clearly distinguished. Refraction and dark-field were found to highlight surface roughness formed during solidification. This work demonstrates that the availability of the additional contrast channels provided by multi-contrast X-ray imaging delivers additional information, also when imaging high atomic number specimens with a significant absorption.

A laboratory-based, low-energy, multi-modal x-ray microscope with user-defined resolution.

We report on the development of a low-energy x-ray phase-based microscope using intensity-modulation masks for single-shot retrieval of three contrast channels: transmission, refraction, and ultra-small-angle scattering or dark field. The retrieval method is based on beam tracking, an incoherent and phase-based imaging approach. We demonstrate that the spatial resolution of this imaging system does not depend on focal spot size nor detector pixel pitch, as opposed to conventional and propagation-based x-ray imaging, and it is only dependent on the mask aperture size. This result enables the development of a multi-resolution microscope where multi-scale samples can be explored on different length scales by adjusting only the mask aperture size, without other modifications. Additionally, we show an extended capability of the system to resolve periodic structures below the resolution limit imposed by the mask apertures, which potentially extends dark-field imaging beyond its conventional use.

Monitoring tissue engineered constructs and protocols with laboratory-based x-ray phase contrast tomography.

Tissue engineering (TE) aims to generate bioengineered constructs which can offer a surgical treatment for many conditions involving tissue or organ loss. Construct generation must be guided by suitable assessment tools. However, most current tools (e.g. histology) are destructive, which restricts evaluation to a single-2D anatomical plane, and has no potential for assessing constructs prior to or following their implantation. An alternative can be provided by laboratory-based x-ray phase contrast computed tomography (PC-CT), which enables the extraction of 3D density maps of an organ's anatomy. In this work, we developed a semi-automated image processing pipeline dedicated to the analysis of PC-CT slices of oesophageal constructs. Visual and quantitative (density and morphological) information is extracted on a volumetric basis, enabling a comprehensive evaluation of the regenerated constructs. We believe the presented tools can enable the successful regeneration of patient-specific oesophagus, and bring comparable benefit to a wide range of TE applications. STATEMENT OF SIGNIFICANCE: Phase contrast computed tomography (PC-CT) is an imaging modality which generates high resolution volumetric density maps of biological tissue. In this work, we demonstrate the use of PC-CT as a new tool for guiding the progression of an oesophageal tissue engineering (TE) protocol. Specifically, we developed a semi-automated image-processing pipeline which analyses the oesophageal PC-CT slices, extracting visual and quantitative (density and morphological) information. This information was proven key for performing a comprehensive evaluation of the regenerated constructs, and cannot be obtained through existing assessment tools primarily due to their destructive nature (e.g. histology). This work paves the way for using PC-CT in a wide range of TE applications which can be pivotal for unlocking the potential of this field.

Volumetric High-Resolution X-Ray Phase-Contrast Virtual Histology of Breast Specimens With a Compact Laboratory System.

The assessment of margin involvement is a fundamental task in breast conserving surgery to prevent recurrences and reoperations. It is usually performed through histology, which makes the process time consuming and can prevent the complete volumetric analysis of large specimens. X-ray phase contrast tomography combines high resolution, sufficient penetration depth and high soft tissue contrast, and can therefore provide a potential solution to this problem. In this work, we used a high-resolution implementation of the edge illumination X-ray phase contrast tomography based on "pixel-skipping" X-ray masks and sample dithering, to provide high definition virtual slices of breast specimens. The scanner was originally designed for intra-operative applications in which short scanning times were prioritised over spatial resolution; however, thanks to the versatility of edge illumination, high-resolution capabilities can be obtained with the same system simply by swapping x-ray masks without this imposing a reduction in the available field of view. This makes possible an improved visibility of fine tissue strands, enabling a direct comparison of selected CT slices with histology, and providing a tool to identify suspect features in large specimens before slicing. Combined with our previous results on fast specimen scanning, this works paves the way for the design of a multi-resolution EI scanner providing intra-operative capabilities as well as serving as a digital pathology system.

Dynamic Multicontrast X-Ray Imaging Method Applied to Additive Manufacturing.

We present a dynamic implementation of the beam-tracking x-ray imaging method providing absorption, phase, and ultrasmall angle scattering signals with microscopic resolution and high frame rate. We demonstrate the method's ability to capture dynamic processes with 22-ms time resolution by investigating the melting of metals in laser additive manufacturing, which has so far been limited to single-modality synchrotron radiography. The simultaneous availability of three contrast channels enables earlier segmentation of droplets, tracking of powder dynamic, and estimation of unfused powder amounts, demonstrating that the method can provide additional information on melting processes.

Optimization of multipoint phase retrieval in edge illumination X-ray imaging: A theoretical and experimental analysis.

PURPOSE: In this work, an analytical model describing the noise in the retrieved three contrast channels, transmission, refraction, and ultra small-angle scattering, obtained with edge illumination X-ray phase-based imaging system is presented and compared to experimental data. METHODS: In EI, images acquired at different displacements of the presample mask (i.e., different illumination levels referred to as points on the "illumination curve"), followed by pixel-wise curve fitting, are exploited to quantitatively retrieve the three contrast channels. Therefore, the noise in the final image will depend on the error associated with the fitting process. We use a model based on the derivation of the standard error on fitted parameters, which relies on the calculation of the covariance matrix, to estimate the noise and the cross-channel correlation as a function of the position of the sampling points. In particular, we investigated the most common cases of 3 and 5 sampling points. In addition, simulations have been used to better understand the role of the integration time for each sampling point. Finally, the model is validated by comparison with the experimental data acquired with an edge illumination setup based on a tungsten rotating anode X-ray source and a photon counting detector. RESULTS: We found a good match between the predictions of the model and the experimental data. In particular, for the investigated cases, an arrangement of the sampling points leading to minimum noise and cross-channel correlation can be found. Simulations revealed that, given a fixed overall scanning time, its distribution into the smallest possible number of sampling points needed for phase retrieval leads to minimum noise thanks to higher statistics per point. CONCLUSIONS: This work presents an analytical model describing the noise in the various contrast channels retrieved in edge illumination as a function of the illumination curve sampling. In particular, an optimal sampling scheme leading to minimum noise has been determined for the case where 3 or 5 sampling points are used, which represent two of the most common acquisition schemes. In addition, the correlation between noise in the different channels and the role of the number of points and exposure time have been also investigated. In general, our results suggest a series of procedures that should be followed in order to optimize the experimental acquisitions.

Technical Note: Practical implementation strategies of cycloidal computed tomography.

PURPOSE: Cycloidal computed tomography is a novel imaging concept which combines a highly structured x-ray beam, offset lateral under-sampling, and mathematical data recovery to obtain high-resolution images efficiently and flexibly, even with relatively large source focal spots and detector pixels. The method reduces scanning time and, potentially, delivered dose compared to other sampling schemes. This study aims to present and discuss several implementation strategies for cycloidal computed tomography (CT) in order to increase its ease of use and facilitate uptake within the imaging community. METHODS: The different implementation strategies presented are step-and-shoot, continuous unidirectional, continuous back-and-forth, and continuous pixel-wise scanning. In step-and-shoot scans the sample remains stationary while frames are acquired, whereas in all other cases the sample moves through the scanner continuously. The difference between the continuous approaches is the trajectory by which the sample moves within the field of view. RESULTS: All four implementation strategies are compatible with a standard table-top x-ray setup. With the experimental setup applied here, step-and-shoot acquisitions yield the best spatial resolution (around 30 µm), but are the most time-consuming (1.4 h). Continuous unidirectional and back-and-forth images have resolution between 30 and 40 µm, and are faster (35 min). Continuous pixel-wise images are equally time-efficient, although technical challenges caused a small loss in image quality with a resolution of about 50 µm. CONCLUSION: The authors show that cycloidal CT can be implemented in a variety of ways with high quality results. They believe this posits cycloidal CT as a powerful imaging alternative to more time-consuming and less flexible methods in the field.

Improved visualization of X-ray phase contrast volumetric data through artifact-free integrated differential images.

Artifacts arising when differential phase images are integrated is a common problem to several X-ray phase-based experimental techniques. The combination of noise and insufficient sampling of the high-frequency differential phase signal leads to the formation of streak artifacts in the projections, translating into poor image quality in the tomography slices. In this work, we apply a non-iterative integration algorithm proven to reduce streak artifacts in planar (2D) images to a differential phase tomography scan. We report on how the reduction of streak artifacts in the projections improves the quality of the tomography slices, especially in the directions different from the reconstruction plane. Importantly, the method is compatible with large tomography datasets in terms of computation time.

Detection of involved margins in breast specimens with X-ray phase-contrast computed tomography.

Margins of wide local excisions in breast conserving surgery are tested through histology, which can delay results by days and lead to second operations. Detection of margin involvement intraoperatively would allow the removal of additional tissue during the same intervention. X-ray phase contrast imaging (XPCI) provides soft tissue sensitivity superior to conventional X-rays: we propose its use to detect margin involvement intraoperatively. We have developed a system that can perform phase-based computed tomography (CT) scans in minutes, used it to image 101 specimens approximately half of which contained neoplastic lesions, and compared results against those of a commercial system. Histological analysis was carried out on all specimens and used as the gold standard. XPCI-CT showed higher sensitivity (83%, 95% CI 69-92%) than conventional specimen imaging (32%, 95% CI 20-49%) for detection of lesions at margin, and comparable specificity (83%, 95% CI 70-92% vs 86%, 95% CI 73-93%). Within the limits of this study, in particular that specimens obtained from surplus tissue typically contain small lesions which makes detection more difficult for both methods, we believe it likely that the observed increase in sensitivity will lead to a comparable reduction in the number of re-operations.

Evaluation of a compact multicontrast and multiresolution X-ray phase contrast edge illumination system for small animal imaging.

PURPOSE: In this work the performance of a compact multiresolution and multicontrast x-ray phase system based on edge illumination is investigated. It has been designed for small animal imaging and with a limited footprint for ease of deployment in laboratories. METHODS: The presented edge illumination system is based on a compact microfocus tungsten x-ray source combined with a flat panel detector. The source has a maximum output of 10 W when the minimum spot size of about 15 µm is used. The system has an overall length of 70 cm. A new double sample mask design, obtained by arranging both skipped and nonskipped configurations on the same structure, provides dual resolution capability. To test the system, we carried out computed tomography (CT) scans of a plastic phantom with different source settings using both single-image and multi-image acquisition schemes at different spatial resolutions. In addition, CT scans of an ex-vivo mouse specimen were acquired at the best identified working conditions to demonstrate the application of the presented system to small animal imaging. RESULTS: We found this system delivers good image quality, allowing for an efficient material separation and improving detail visibility in small animals thanks to the higher signal-to-noise ratio (SNR) of phase contrast with respect to conventional attenuation contrast. The system offers high versatility in terms of spatial resolution thanks to the double sample mask design integrated into a single scanner. The availability of both multi- and single-image acquisition schemes coupled with their dedicated retrieval algorithms, allows different working modes which can be selected based on user preference. Multi-image acquisition provides quantitative separation of the real and imaginary part of the refractive index, however, it requires a long scanning time. On the other hand, the single image approach delivers the best material separation and image quality at all the investigated source settings with a shorter scanning time but at the cost of quantitativeness. Finally, we also observed that the single image approach combined with a high-power x-ray source may result in a fast acquisition protocol compatible with in-vivo imaging.

Fast, non-iterative algorithm for quantitative integration of X-ray differential phase-contrast images.

X-ray phase contrast imaging is gaining importance as an imaging tool. However, it is common for X-ray phase detection techniques to be sensitive to the derivatives of the phase. Therefore, the integration of differential phase images is a fundamental step both to access quantitative pixel content and for further analysis such as segmentation. The integration of noisy data leads to artefacts with a severe impact on image quality and on its quantitative content. In this work, an integration method based on the Wiener filter is presented and tested using simulated and real data obtained with the edge illumination differential X-ray phase imaging method. The method is shown to provide high image quality while preserving the quantitative pixel content of the integrated image. In addition, it requires a short computational time making it suitable for large datasets.

Simplified retrieval method for Edge Illumination X-ray phase contrast imaging allowing multi-modal imaging with fewer input frames.

We present data from an implementation of Edge Illumination (EI) that uses a detector aperture designed for increasing dynamic range, suitable for clinically relevant X-ray energies and demonstrated here using synchrotron radiation. By utilising a sufficiently large crosstalk between pixels, this implementation enables single-scan imaging for phase and absorption, and double-scan for phase, absorption and dark field imaging. The presence of the detector mask enables a direct comparison between conventional EI and beam tracking (BT), which we conduct through Monte Carlo and analytical modelling in the case of a single-scan being used for the retrieval of all three contrasts. In the present case, where the X-ray beam width is comparable to the pixel size, we provide an analysis on best-positioning of the beam on the detector for accurate signal retrieval. Further, we demonstrate an application of this method by distinguishing different concentrations of microbubbles via their dark field signals at high energy using an EI system.

A Preliminary Investigation into the Use of Edge Illumination X-ray Phase Contrast Micro-CT for Preclinical Imaging.

PURPOSE: To enable a preliminary assessment of the suitability of edge illumination (EI) x-ray phase contrast (XPC) micro x-ray computed tomography (micro-CT) to preclinical imaging. Specifically, to understand how different acquisition schemes and their combination with dedicated data processing affect contrast-to-noise ratio (CNR) and spatial resolution, while providing control over scan time and radiation dose delivery. PROCEDURES: Deceased mice (n = 3) were scanned with an EI XPC micro-CT setup operated under different settings, leading to scan times between 18 h and 13 min. For the shortest scan, the entrance dose was measured with a calibrated PTW 23344 ion chamber. Different data processing methods were applied, retrieving either separate attenuation and phase images, or hybrid (combined attenuation and phase) images. A quantitative comparison was performed based on CNR and spatial resolution measurements for a soft tissue interface. RESULTS: All phase-based images have led to a higher CNR for the considered soft tissue interface than the attenuation image, independent of scan time. The best relative CNR (a sixfold increase) was observed in one of the hybrid images. Spatial resolution was found to be connected to scan time, with a resolution of approximately 20 µm and 60 µm achieved for the longest and shortest scans, respectively. An entrance dose of approximately 300 mGy was estimated for the scan performed within 13 min. CONCLUSIONS: Despite their preliminary nature, our results suggest that EI XPC bears potential for enhancing the utility of preclinical micro-CT, and, pending further research and development, could ultimately become a valuable technique in this field.