A feasibility study of X‐ray phase‐contrast mammographic tomography at the Imaging and Medical beamline of the Australian Synchrotron

Results are presented of a recent experiment at the Imaging and Medical beamline of the Australian Synchrotron intended to contribute to the implementation of low‐dose high‐sensitivity three‐dimensional mammographic phase‐contrast imaging, initially at synchrotrons and subsequently in hospitals and medical imaging clinics. The effect of such imaging parameters as X‐ray energy, source size, detector resolution, sample‐to‐detector distance, scanning and data processing strategies in the case of propagation‐based phase‐contrast computed tomography (CT) have been tested, quantified, evaluated and optimized using a plastic phantom simulating relevant breast‐tissue characteristics. Analysis of the data collected using a Hamamatsu CMOS Flat Panel Sensor, with a pixel size of 100 µm, revealed the presence of propagation‐based phase contrast and demonstrated significant improvement of the quality of phase‐contrast CT imaging compared with conventional (absorption‐based) CT, at medically acceptable radiation doses.     

Hard X‐ray phase‐contrast tomography of non‐homogeneous specimens: grating interferometry versus propagation‐based imaging

X‐ray phase‐contrast imaging is an effective approach to drastically increase the contrast and sensitivity of microtomographic techniques. Numerous approaches to depict the real part of the complex‐valued refractive index of a specimen are nowadays available. A comparative study using experimental data from grating‐based interferometry and propagation‐based phase contrast combined with single‐distance phase retrieval applied to a non‐homogeneous sample is presented (acquired at beamline ID19‐ESRF). It is shown that grating‐based interferometry can handle density gradients in a superior manner. The study underlines the complementarity of the two techniques for practical applications.     

Advanced phase‐contrast imaging using a grating interferometer

Phase‐sensitive X‐ray imaging methods can provide substantially increased contrast over conventional absorption‐based imaging, and therefore new and otherwise inaccessible information. Differential phase‐contrast (DPC) imaging, which uses a grating interferometer and a phase‐stepping technique, has been integrated into TOMCAT, a beamline dedicated to tomographic microscopy and coherent radiology experiments at the Swiss Light Source. Developments have been made focusing on the fast acquisition and post‐processing of data to enable a high‐throughput of samples, with obvious advantages, also through increasing the efficiency of the detecting system, of helping to reduce radiation dose imparted to the sample. A novel aquarium design allows a vertical rotation axis below the sample with measurements performed in aqueous environment. Optimization of the data acquisition procedure enables a full phase volume (1024 × 1024 pixels × 1000 projections × 9 phase steps, i.e. 9000 projections in total) to be acquired in 20 min (with a pixel size of 7.4 µm), and the subsequent post‐processing has been integrated into the beamline pipeline for sinogram generation. Local DPC tomography allows one to focus with higher magnification on a particular region of interest of a sample without the presence of local tomography reconstruction artifacts. Furthermore, `widefield' imaging is shown for DPC scans for the first time, enabling the field of view of the imaging system to be doubled for samples that are larger than the magnification allows. A case study is illustrated focusing on the visualization of soft tissue features, and particularly the substantia nigra of a rat brain. Darkfield images, based on local X‐ray scattering, can also be extracted from a grating‐based DPC scan: an example of the advantages of darkfield contrast is shown and the potential of darkfield X‐ray tomography is discussed.     

X‐ray grating interferometer for biomedical imaging applications at Shanghai Synchrotron Radiation Facility

An X‐ray grating interferometer was installed at the BL13W beamline of Shanghai Synchrotron Radiation Facility (SSRF) for biomedical imaging applications. Compared with imaging results from conventional absorption‐based micro‐computed tomography, this set‐up has shown much better soft tissue imaging capability. In particular, using the set‐up, the carotid artery and the carotid vein in a formalin‐fixed mouse can be visualized in situ without contrast agents, paving the way for future applications in cancer angiography studies. The overall results have demonstrated the broad prospects of the existing set‐up for biomedical imaging applications at SSRF.     

Feasibility study of propagation‐based phase‐contrast X‐ray lung imaging on the Imaging and Medical beamline at the Australian Synchrotron

Propagation‐based phase‐contrast X‐ray imaging (PB‐PCXI) using synchrotron radiation has achieved high‐resolution imaging of the lungs of small animals both in real time and in vivo. Current studies are applying such imaging techniques to lung disease models to aid in diagnosis and treatment development. At the Australian Synchrotron, the Imaging and Medical beamline (IMBL) is well equipped for PB‐PCXI, combining high flux and coherence with a beam size sufficient to image large animals, such as sheep, due to a wiggler source and source‐to‐sample distances of over 137 m. This study aimed to measure the capabilities of PB‐PCXI on IMBL for imaging small animal lungs to study lung disease. The feasibility of combining this technique with computed tomography for three‐dimensional imaging and X‐ray velocimetry for studies of airflow and non‐invasive lung function testing was also investigated. Detailed analysis of the role of the effective source size and sample‐to‐detector distance on lung image contrast was undertaken as well as phase retrieval for sample volume analysis. Results showed that PB‐PCXI of lung phantoms and mouse lungs produced high‐contrast images, with successful computed tomography and velocimetry also being carried out, suggesting that live animal lung imaging will also be feasible at the IMBL.     

Potential of propagation‐based synchrotron X‐ray phase‐contrast computed tomography for cardiac tissue engineering

Hydrogel‐based cardiac tissue engineering offers great promise for myocardial infarction repair. The ability to visualize engineered systems in vivo in animal models is desired to monitor the performance of cardiac constructs. However, due to the low density and weak X‐ray attenuation of hydrogels, conventional radiography and micro‐computed tomography are unable to visualize the hydrogel cardiac constructs upon their implantation, thus limiting their use in animal systems. This paper presents a study on the optimization of synchrotron X‐ray propagation‐based phase‐contrast imaging computed tomography (PCI‐CT) for three‐dimensional (3D) visualization and assessment of the hydrogel cardiac patches. First, alginate hydrogel was 3D‐printed into cardiac patches, with the pores filled by fibrin. The hydrogel patches were then surgically implanted on rat hearts. A week after surgery, the hearts including patches were excised and embedded in a soft‐tissue‐mimicking gel for imaging by using PCI‐CT at an X‐ray energy of 25 keV. During imaging, the sample‐to‐detector distances, CT‐scan time and the region of interest (ROI) were varied and examined for their effects on both imaging quality and radiation dose. The results showed that phase‐retrieved PCI‐CT images provided edge‐enhancement fringes at a sample‐to‐detector distance of 147 cm that enabled visualization of anatomical and microstructural features of the myocardium and the implanted patch in the tissue‐mimicking gel. For visualization of these features, PCI‐CT offered a significantly higher performance than the dual absorption‐phase and clinical magnetic resonance (3 T) imaging techniques. Furthermore, by reducing the total CT‐scan time and ROI, PCI‐CT was examined for lowering the effective dose, meanwhile without much loss of imaging quality. In effect, the higher soft tissue contrast and low‐dose potential of PCI‐CT has been used along with an acceptable overall animal dose to achieve the high spatial resolution needed for cardiac implant visualization. As a result, PCI‐CT at the identified imaging parameters offers great potential for 3D assessment of microstructural features of hydrogel cardiac patches.     

First experiments on the Australian Synchrotron Imaging and Medical beamline,including investigations of the effective source size in respect of X‐ray imaging

The Imaging and Medical beamline at the Australian Synchrotron achieved `first light' in December 2008. Here, the first experiments performed on the beamline are reported, which involved both X‐ray imaging and tomography studies for a range of samples. The use of a plastic‐edge phantom for quantitative measurements of contrast and resolution proved to be very instructive and helped to confirm certain parameter values such as the effective horizontal source size, detector resolution and average X‐ray energy for the polychromatic beam.     

X‐ray analyzer‐based phase‐contrast computed laminography

X‐ray analyzer‐based phase‐contrast imaging is combined with computed laminography for imaging regions of interest in laterally extended flat specimens of weak absorption contrast. The optics discussed here consist of an asymmetrically cut collimator crystal and a symmetrically cut analyzer crystal arranged in a nondispersive (+, ?) diffraction geometry. A generalized algorithm is given for calculating multi‐contrast (absorption, refraction and phase contrast) images of a sample. Basic formulae are also presented for laminographic reconstruction. The feasibility of the method discussed was verified at the vertical wiggler beamline BL‐14B of the Photon Factory. At a wavelength of 0.0733 nm, phase‐contrast sectional images of plastic beads were successfully obtained. Owing to strong circular artifacts caused by a sample holder, the field of view was limited to about 6 mm in diameter.     

MISTRAL: a transmission soft X‐ray microscopy beamline for cryo nano‐tomography of biological samples and magnetic domains imaging

The performance of MISTRAL is reported, the soft X‐ray transmission microscopy beamline at the ALBA light source (Barcelona, Spain) which is primarily dedicated to cryo soft X‐ray tomography (cryo‐SXT) for three‐dimensional visualization of whole unstained cells at spatial resolutions down to 30 nm (half pitch). Short acquisition times allowing for high‐throughput and correlative microscopy studies have promoted cryo‐SXT as an emerging cellular imaging tool for structural cell biologists bridging the gap between optical and electron microscopy. In addition, the beamline offers the possibility of imaging magnetic domains in thin magnetic films that are illustrated here with an example.     

PITRE: software for phase‐sensitive X‐ray image processing and tomography reconstruction

Synchrotron‐radiation computed tomography has been applied in many research fields. Here, PITRE (Phase‐sensitive X‐ray Image processing and Tomography REconstruction) and PITRE_BM (PITRE Batch Manager) are presented. PITRE supports phase retrieval for propagation‐based phase‐contrast imaging/tomography (PPCI/PPCT), extracts apparent absorption, refractive and scattering information of diffraction enhanced imaging (DEI), and allows parallel‐beam tomography reconstruction for conventional absorption CT data and for PPCT phase retrieved and DEI‐CT extracted information. PITRE_BM is a batch processing manager for PITRE: it executes a series of tasks, created viaPITRE, without manual intervention. Both PITRE and PITRE_BM are coded in Interactive Data Language (IDL), and have a user‐friendly graphical user interface. They are freeware and can run on Microsoft Windows systems via IDL Virtual Machine, which can be downloaded for free and does not require a license. The data‐processing principle and some examples of application will be presented.     

White‐beam X‐ray radioscopy and tomography with simultaneous diffraction at the EDDI beamline

A set‐up for simultaneous imaging and diffraction that yields radiograms with up to 200 frames per second and 5.6 µm effective pixel size is presented. Tomograms and diffractograms are acquired together in 10 s. Two examples illustrate the attractiveness of combining these methods at the EDDI beamline for in situ studies.     

Comparison of in vitro breast cancer visibility in analyser‐based computed tomography with histopathology,mammography, computed tomography and magnetic resonance imaging

High‐resolution analyser‐based X‐ray imaging computed tomography (HR ABI‐CT) findings on in vitro human breast cancer are compared with histopathology, mammography, computed tomography (CT) and magnetic resonance imaging. The HR ABI‐CT images provided significantly better low‐contrast visibility compared with the standard radiological images. Fine cancer structures indistinguishable and superimposed in mammograms were seen, and could be matched with the histopathological results. The mean glandular dose was less than 1 mGy in mammography and 12–13 mGy in CT and ABI‐CT. The excellent visibility of in vitro breast cancer suggests that HR ABI‐CT may have a valuable role in the future as an adjunct or even alternative to current breast diagnostics, when radiation dose is further decreased, and compact synchrotron radiation sources become available.     

Analysis and interpretation of the first monochromatic X‐ray tomography data collected at the Australian Synchrotron Imaging and Medical beamline

The first monochromatic X‐ray tomography experiments conducted at the Imaging and Medical beamline of the Australian Synchrotron are reported. The sample was a phantom comprising nylon line, Al wire and finer Cu wire twisted together. Data sets were collected at four different X‐ray energies. In order to quantitatively account for the experimental values obtained for the Hounsfield (or CT) number, it was necessary to consider various issues including the point‐spread function for the X‐ray imaging system and harmonic contamination of the X‐ray beam. The analysis and interpretation of the data includes detailed considerations of the resolution and efficiency of the CCD detector, calculations of the X‐ray spectrum prior to monochromatization, allowance for the response of the double‐crystal Si monochromator used (via X‐ray dynamical theory), as well as a thorough assessment of the role of X‐ray phase‐contrast effects. Computer simulations relating to the tomography experiments also provide valuable insights into these important issues. It was found that a significant discrepancy between theory and experiment for the Cu wire could be largely resolved in terms of the effect of the point‐spread function. The findings of this study are important in respect of any attempts to extract quantitative information from X‐ray tomography data, across a wide range of disciplines, including materials and life sciences.     

Live small‐animal X‐ray lung velocimetry and lung micro‐tomography at the Australian Synchrotron Imaging and Medical Beamline

The high flux and coherence produced at long synchrotron beamlines makes them well suited to performing phase‐contrast X‐ray imaging of the airways and lungs of live small animals. Here, findings of the first live‐animal imaging on the Imaging and Medical Beamline (IMBL) at the Australian Synchrotron are reported, demonstrating the feasibility of performing dynamic lung motion measurement and high‐resolution micro‐tomography. Live anaesthetized mice were imaged using 30 keV monochromatic X‐rays at a range of sample‐to‐detector propagation distances. A frame rate of 100 frames s^?1 allowed lung motion to be determined using X‐ray velocimetry. A separate group of humanely killed mice and rats were imaged by computed tomography at high resolution. Images were reconstructed and rendered to demonstrate the capacity for detailed, user‐directed display of relevant respiratory anatomy. The ability to perform X‐ray velocimetry on live mice at the IMBL was successfully demonstrated. High‐quality renderings of the head and lungs visualized both large structures and fine details of the nasal and respiratory anatomy. The effect of sample‐to‐detector propagation distance on contrast and resolution was also investigated, demonstrating that soft tissue contrast increases, and resolution decreases, with increasing propagation distance. This new capability to perform live‐animal imaging and high‐resolution micro‐tomography at the IMBL enhances the capability for investigation of respiratory diseases and the acceleration of treatment development in Australia.     

Design of a mouse restraint for synchrotron‐based computed tomography imaging

High‐resolution computed tomography (CT) imaging of a live animal within a lead‐lined synchrotron light hutch presents several unique challenges. In order to confirm that the animal is under a stable plane of anaesthesia, several physiological parameters (e.g. heart rate, arterial oxygen saturation, core body temperature and respiratory rate) must be remotely monitored from outside the imaging hutch. In addition, to properly scan the thoracic region using CT, the animal needs to be held in a vertical position perpendicular to the fixed angle of the X‐ray beam and free to rotate 180°–360°. A new X‐ray transparent mouse restraint designed and fabricated using computer‐aided design software and three‐dimensional rapid prototype printing has been successfully tested at the Biomedical Imaging and Therapy bending‐magnet (BMIT‐BM) beamline at the Canadian Light Source.     

Characterization of a sCMOS‐based high‐resolution imaging system

The detection system is a key part of any imaging station. Here the performance of the novel sCMOS‐based detection system installed at the ID17 biomedical beamline of the European Synchrotron Radiation Facility and dedicated to high‐resolution computed‐tomography imaging is analysed. The system consists of an X‐ray–visible‐light converter, a visible‐light optics and a PCO.Edge5.5 sCMOS detector. Measurements of the optical characteristics, the linearity of the system, the detection lag, the modulation transfer function, the normalized power spectrum, the detective quantum efficiency and the photon transfer curve are presented and discussed. The study was carried out at two different X‐ray energies (35 and 50 keV) using both 2× and 1× optical magnification systems. The final pixel size resulted in 3.1 and 6.2 µm, respectively. The measured characteristic parameters of the PCO.Edge5.5 are in good agreement with the manufacturer specifications. Fast imaging can be achieved using this detection system, but at the price of unavoidable losses in terms of image quality. The way in which the X‐ray beam inhomogeneity limited some of the performances of the system is also discussed.     

Early commissioning results for spectroscopic X‐ray Nano‐Imaging Beamline BL 7C sXNI at PLS‐II

For spectral imaging of chemical distributions using X‐ray absorption near‐edge structure (XANES) spectra, a modified double‐crystal monochromator, a focusing plane mirrors system and a newly developed fluorescence‐type X‐ray beam‐position monitoring and feedback system have been implemented. This major hardware upgrade provides a sufficiently stable X‐ray source during energy scanning of more than hundreds of eV for acquisition of reliable XANES spectra in two‐dimensional and three‐dimensional images. In recent pilot studies discussed in this paper, heavy‐metal uptake by plant roots in vivo and iron's phase distribution in the lithium–iron–phosphate cathode of a lithium‐ion battery have been imaged. Also, the spatial resolution of computed tomography has been improved from 70 nm to 55 nm by means of run‐out correction and application of a reconstruction algorithm.     

Phase‐space analysis and experimental results for secondary focusing at X‐ray beamlines

Micro‐focusing optical devices at synchrotron beamlines usually have a limited acceptance, but more flux can be intercepted if such optics are used to focus secondary sources created by the primary optics. Flux throughput can be maximized by placing the secondary focusing optics close to or exactly at the secondary source position. However, standard methods of beamline optics analysis, such as the lens equation or matching the mirror surface to an ellipse, work poorly when the source‐to‐optics distance is very short. In this paper the general characteristics of the focusing of beams with Gaussian profiles by a `thin lens' are analysed under the paraxial approximation in phase space, concluding that the focusing of a beam with a short source‐to‐optics distance is distinct from imaging the source; slope errors are successfully included in all the formulas so that they can be used to calculate beamline focusing with good accuracy. A method is also introduced to use the thin‐lens result to analyse the micro‐focusing produced by an elliptically bent trapezoid‐shaped Kirkpatrick–Baez mirror. The results of this analysis are in good agreement with ray‐tracing simulations and are confirmed by the experimental results of the secondary focusing at the 18‐ID Bio‐CAT beamline (at the APS). The result of secondary focusing carried out at 18‐ID using a single‐bounce capillary can also be explained using this phase‐space analysis. A discussion of the secondary focusing results is presented at the end of this paper.     

Analyser‐based tomography images of cartilage

Analyser‐based imaging expands the performance of X‐ray imaging by utilizing not only the absorption properties of X‐rays but also the refraction and scatter rejection (extinction) properties. In this study, analyser‐based computed tomography has been implemented on imaging an articular cartilage sample, depicting substructural variations, without overlay, at a pixel resolution of 3.6 µm.     

In vivo physiological saline‐infused hepatic vessel imaging using a two‐crystal‐interferometer‐based phase‐contrast X‐ray technique

Using a two‐crystal‐interferometer‐based phase‐contrast X‐ray imaging system, the portal vein, capillary vessel area and hepatic vein of live rats were revealed sequentially by injecting physiological saline via the portal vein. Vessels greater than 0.06 mm in diameter were clearly shown with low levels of X‐rays (552 µGy). This suggests that in vivo vessel imaging of small animals can be performed as conventional angiography without the side effects of the presently used iodine contrast agents.