Dry deposition of pollutant and marker particles onto live mouse airway surfaces enhances monitoring of individual particle mucociliary transit behaviour

Particles suspended in the air are inhaled during normal respiration and unless cleared by airway defences, such as the mucociliary transit (MCT) system, they can remain and affect lung and airway health. Synchrotron phase‐contrast X‐ray imaging (PCXI) methods have been developed to non‐invasively monitor the behaviour of individual particles in live mouse airways and in previous studies the MCT behaviour of particles and fibres in the airways of live mice after deposition in a saline carrier fluid have been examined. In this study a range of common respirable pollutant particles (lead dust, quarry dust and fibreglass fibres) as well as marker particles (hollow glass micro‐spheres) were delivered into the trachea of live mice using a dry powder insufflator to more accurately mimic normal environmental particulate exposure and deposition via inhalation. The behaviour of the particles once delivered onto the airway surface was tracked over a five minute period via PCXI. All particles were visible after deposition. Fibreglass fibres remained stationary throughout while all other particle types transited the tracheal surface throughout the imaging period. In all cases the majority of the particle deposition and any airway surface activity was located close to the dorsal tracheal wall. Both the individual and bulk motions of the glass bead marker particles were visible and their behaviour enabled otherwise hidden MCT patterns to be revealed. This study verified the value of PCXI for examining the post‐deposition particulate MCT behaviour in the mouse trachea and highlighted that MCT is not a uniform process as suggested by radiolabel studies. It also directly revealed the advantages of dry particle delivery for establishing adequate particulate presence for visualizing MCT behaviour. The MCT behaviour and rate seen after dry particle delivery was different from that in previous carrier‐fluid studies. It is proposed that dry particle delivery is essential for producing environmentally realistic particle deposition and studying how living airway surfaces handle different types of inhaled particles by MCT processes.     

A new technique to examine individual pollutant particle and fibre deposition and transit behaviour in live mouse trachea

During respiration, particles suspended in the air are inhaled and unless cleared by airway defences they can remain and affect lung health. Their size precludes the use of standard imaging modalities so we have developed synchrotron phase-contrast X-ray imaging (PCXI) methods to non-invasively monitor the behaviour of individual particles in live mouse airways. In this study we used these techniques to examine post-deposition particle behaviour in the trachea. PCXI was used to monitor the deposition and subsequent behaviour of particles of quarry dust and lead ore; fibres of asbestos and fibreglass; and hollow glass micro-spheres. Visibility was examined in vitro and ex vivo to avoid the complicating effects of surrounding tissue and respiratory or cardiac motion. Particle behaviour was then examined after deposition onto the tracheal airway surfaces of live mice. Each particle and fibre looked and behaved differently on the airway surface. Particles lodged on the airway shortly after deposition, and the rate at which this occurred was dependent on the particle type and size. After the live-imaging experiments, excised airway samples were examined using light and electron microscopy. Evidence of particle capture into the airway surface fluids and the epithelial cell layer was found. PCXI is a valuable tool for examining post-deposition particulate behaviour in the tracheal airway. These first indications that the interaction between airways and individual particles may depend on the particle type and size should provide a novel approach to studying the early effects of respired particles on airway health.     

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.     

Tracking extended mucociliary transport activity of individual deposited particles: longitudinal synchrotron X‐ray imaging in live mice

To assess potential therapies for respiratory diseases in which mucociliary transit (MCT) is impaired, such as cystic fibrosis and primary ciliary dyskinesia, a novel and non‐invasive MCT quantification method has been developed in which the transit rate and behaviour of individual micrometre‐sized deposited particles are measured in live mice using synchrotron phase‐contrast X‐ray imaging. Particle clearance by MCT is known to be a two‐phase process that occurs over a period of minutes to days. Previous studies have assessed MCT in the fast‐clearance phase, ～20 min after marker particle dosing. The aim of this study was to non‐invasively image changes in particle presence and MCT during the slow‐clearance phase, and simultaneously determine whether repeat synchrotron X‐ray imaging of mice was feasible over periods of 3, 9 and 25 h. All mice tolerated the repeat imaging procedure with no adverse effects. Quantitative image analysis revealed that the particle MCT rate and the number of particles present in the airway both decreased with time. This study successfully demonstrated for the first time that longitudinal synchrotron X‐ray imaging studies are possible in live small animals, provided appropriate animal handling techniques are used and care is taken to reduce the delivered radiation dose.     

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.     

Multiple‐element spectrometer for non‐resonant inelastic X‐ray spectroscopy of electronic excitations

A multiple‐analyser‐crystal spectrometer for non‐resonant inelastic X‐ray scattering spectroscopy installed at beamline ID16 of the European Synchrotron Radiation Facility is presented. Nine analyser crystals with bending radii R = 1 m measure spectra for five different momentum transfer values simultaneously. Using a two‐dimensional detector, the spectra given by all analysers can be treated individually. The spectrometer is based on a Rowland circle design with fixed Bragg angles of about 88°. The energy resolution can be chosen between 30–2000 meV with typical incident‐photon energies of 6–13 keV. The spectrometer is optimized for studies of valence and core electron excitations resolving both energy and momentum transfer.     

Synchrotron‐based spectroscopy of X‐ray channeling through hollow capillary microchannels inside glass plates

Here, soft X‐ray synchrotron radiation transmitted through microchannel plates is studied experimentally. Fine structures of reflection and XANES Si L‐edge spectra detected on the exit of silicon glass microcapillary structures under conditions of total X‐ray reflection are presented and analyzed. The phenomenon of the interaction of channeling radiation with unoccupied electronic states and propagation of X‐ray fluorescence excited in the microchannels is revealed. Investigations of the interaction of monochromatic radiation with the inner‐shell capillary surface and propagation of fluorescence radiation through hollow glass capillary waveguides contribute to the development of novel X‐ray focusing devices in the future.     

Carbon contamination of soft X‐ray beamlines: dramatic anti‐reflection coating effects observed in the 1 keV photon energy region

Carbon contamination is a general problem of under‐vacuum optics submitted to high fluence. In soft X‐ray beamlines carbon deposit on optics is known to absorb and scatter radiation close to the C K‐edge (280 eV), forbidding effective measurements in this spectral region. Here the observation of strong reflectivity losses is reported related to carbon deposition at much higher energies around 1000 eV, where carbon absorptivity is small. It is shown that the observed effect can be modelled as a destructive interference from a homogeneous carbon thin film.     

Utilizing broadband X‐rays in a Bragg coherent X‐ray diffraction imaging experiment

A method is presented to simplify Bragg coherent X‐ray diffraction imaging studies of complex heterogeneous crystalline materials with a two‐stage screening/imaging process that utilizes polychromatic and monochromatic coherent X‐rays and is compatible with in situ sample environments. Coherent white‐beam diffraction is used to identify an individual crystal particle or grain that displays desired properties within a larger population. A three‐dimensional reciprocal‐space map suitable for diffraction imaging is then measured for the Bragg peak of interest using a monochromatic beam energy scan that requires no sample motion, thus simplifying in situ chamber design. This approach was demonstrated with Au nanoparticles and will enable, for example, individual grains in a polycrystalline material of specific orientation to be selected, then imaged in three dimensions while under load.     

An electrochemical cell with sapphire windows for operando synchrotron X‐ray powder diffraction and spectroscopy studies of high‐power and high‐voltage electrodes for metal‐ion batteries

A new multi‐purpose operando electrochemical cell was designed, constructed and tested on the Swiss–Norwegian Beamlines BM01 and BM31 at the European Synchrotron Radiation Facility. Single‐crystal sapphire X‐ray windows provide a good signal‐to‐noise ratio, excellent electrochemical contact because of the constant pressure between the electrodes, and perfect electrochemical stability at high potentials due to the inert and non‐conductive nature of sapphire. Examination of the phase transformations in the Li_1–xFe_0.5Mn_0.5PO₄ positive electrode (cathode) material at C/2 and 10C charge and discharge rates, and a study of the valence state of the Ni cations in the Li_1–xNi_0.5Mn_1.5O₄ cathode material for Li‐ion batteries, revealed the applicability of this novel cell design to diffraction and spectroscopic investigations of high‐power/high‐voltage electrodes for metal‐ion batteries.     

The materials science X‐ray beamline BL8 at the DELTA storage ring

The hard X‐ray beamline BL8 at the superconducting asymmetric wiggler at the 1.5 GeV Dortmund Electron Accelerator DELTA is described. This beamline is dedicated to X‐ray studies in the spectral range from ～1 keV to ～25 keV photon energy. The monochromator as well as the other optical components of the beamline are optimized accordingly. The endstation comprises a six‐axis diffractometer that is capable of carrying heavy loads related to non‐ambient sample environments such as, for example, ultrahigh‐vacuum systems, high‐pressure cells or liquid‐helium cryostats. X‐ray absorption spectra from several reference compounds illustrate the performance. Besides transmission measurements, fluorescence detection for dilute sample systems as well as surface‐sensitive reflection‐mode experiments have been performed. The results show that high‐quality EXAFS data can be obtained in the quick‐scanning EXAFS mode within a few seconds of acquisition time, enabling time‐resolved in situ experiments using standard beamline equipment that is permanently available. The performance of the new beamline, especially in terms of the photon flux and energy resolution, is competitive with other insertion‐device beamlines worldwide, and several sophisticated experiments including surface‐sensitive EXAFS experiments are feasible.     

Diffractive–refractive optics: X‐ray splitter

The possibility of splitting a thin (e.g. undulator) X‐ray beam based on diffraction–refraction effects is discussed. The beam is diffracted from a crystal whose diffracting surface has the shape of a roof with the ridge lying in the plane of diffraction. The crystal is cut asymmetrically. One half of the beam impinges on the left‐hand part of the roof and the other half impinges on the right‐hand side of the roof. Owing to refraction the left part of the beam is deviated to the left whereas the right part is deviated to the right. The device proposed consists of two channel‐cut crystals with roof‐like diffraction surfaces; the crystals are set in a dispersive position. The separation of the beams after splitting is calculated at a distance of 10 m from the crystals for various asymmetry and inclination angles. It is shown that such a splitting may be utilized for long beamlines. Advantages and disadvantages of this method are discussed.     

A microfocus X‐ray fluorescence beamline at Indus‐2 synchrotron radiation facility

A microfocus X‐ray fluorescence spectroscopy beamline (BL‐16) at the Indian synchrotron radiation facility Indus‐2 has been constructed with an experimental emphasis on environmental, archaeological, biomedical and material science applications involving heavy metal speciation and their localization. The beamline offers a combination of different analytical probes, e.g. X‐ray fluorescence mapping, X‐ray microspectroscopy and total‐external‐reflection fluorescence characterization. The beamline is installed on a bending‐magnet source with a working X‐ray energy range of 4–20 keV, enabling it to excite K‐edges of all elements from S to Nb and L‐edges from Ag to U. The optics of the beamline comprises of a double‐crystal monochromator with Si(111) symmetric and asymmetric crystals and a pair of Kirkpatrick–Baez focusing mirrors. This paper describes the performance of the beamline and its capabilities with examples of measured results.     

Ultra‐short and ultra‐intense X‐ray free‐electron laser single pulse in one‐dimensional photonic crystals

The propagation within a one‐dimensional photonic crystal of a single ultra‐short and ultra‐intense pulse delivered by an X‐ray free‐electron laser is analysed with the framework of the time‐dependent coupled‐wave theory in non‐linear media. It is shown that the reflection and the transmission of an ultra‐short pulse present a transient period conditioned by the extinction length and also the thickness of the structure for transmission. For ultra‐intense pulses, non‐linear effects are expected: they could give rise to numerous phenomena, bi‐stability, self‐induced transparency, gap solitons, switching, etc., which have been previously shown in the optical domain.     

X‐ray‐Raman‐scattering‐based EXAFS beyond the dipole limit

X‐ray Raman scattering (XRS) provides a bulk‐sensitive method of measuring the extended X‐ray absorption fine structure (EXAFS) of soft X‐ray absorption edges. Accurate measurements and data analysis procedures for the determination of XRS‐EXAFS of polycrystalline diamond are described. The contributions of various angular‐momentum components beyond the dipole limit to the atomic background and the EXAFS oscillations are incorporated using self‐consistent real‐space multiple‐scattering calculations. The properly extracted XRS‐EXAFS oscillations are in good agreement with calculations and earlier soft X‐ray EXAFS results. It is shown, however, that under certain conditions multiple‐scattering contributions to XRS‐EXAFS deviate from those in standard EXAFS, leading to noticeable changes in the real‐space signal at higher momentum transfers owing to non‐dipole contributions. These results pave the way for the accurate application of XRS‐EXAFS to previously inaccessible light‐element systems.     

X‐ray fluorescence induced by standing waves in the grazing‐incidence and grazing‐exit modes: study of the Mg–Co–Zr system

The characterization of Mg–Co–Zr tri‐layer stacks using X‐ray fluorescence induced by X‐ray standing waves, in both the grazing‐incidence (GI) and the grazing‐exit (GE) modes, is presented. The introduction of a slit in the direction of the detector improves the angular resolution by a factor of two and significantly improves the sensitivity of the technique for the chemical characterization of the buried interfaces. By observing the intensity variations of the Mg Kα and Co Lα characteristic emissions as a function of the incident (GI mode) or detection (GE mode) angle, it is shown that the interfaces of the Si/[Mg/Co/Zr]_×30 multilayer are abrupt, whereas in the Si/[Mg/Zr/Co]_×30 multilayer a strong intermixing occurs at the Co‐on‐Zr interfaces. An explanation of this opposite behavior of the Co‐on‐Zr and Zr‐on‐Co interfaces is given by the calculation of the mixing enthalpies of the Co–Mg, Co–Zr and Mg–Zr systems, which shows that the Co–Zr system presents a negative value and the other two systems present positive values. Together with the difference of the surface free energies of Zr and Co, this leads to the Mg/Zr/Co system being considered as a Mg/Co_xZr_y bi‐layer stack, with x/y estimated around 3.5.     

X‐ray photon correlation spectroscopy in systems without long‐range order: existence of an intermediate‐field regime

Successful X‐ray photon correlation spectroscopy studies often require that signals be optimized while minimizing power density in the sample to decrease radiation damage and, at free‐electron laser sources, thermal impact. This suggests exploration of scattering outside the Fraunhofer far‐field diffraction limit d²/λR, where d is the incident beam size, λ is the photon wavelength and R is the sample‐to‐detector distance. Here it is shown that, in an intermediate regime d²/λ > Rdξ/λ, where ξ is the structural correlation length in the material, the ensemble averages of the scattered intensity and of the structure factor are equal. Similarly, in the regime d²/λ > Rdξ(τ)/λ, where ξ(τ) is a time‐dependent dynamics length scale of interest, the ensemble‐averaged correlation functions g₁(τ) and g₂(τ) of the scattered electric field are also equal to their values in the far‐field limit. This broadens the parameter space for X‐ray photon correlation spectroscopy experiments, but detectors with smaller pixel size and variable focusing are required to more fully exploit the potential for such studies.     

X‐ray photon correlation spectroscopy under flow

X‐ray photon correlation spectroscopy was used to probe the diffusive dynamics of colloidal particles in a shear flow. Combining X‐ray techniques with microfluidics is an experimental strategy that reduces the risk of X‐ray‐induced beam damage and also allows time‐resolved studies of processes taking place in flow cells. The experimental results and theoretical predictions presented here show that in the low shear limit for a `transverse flow' scattering geometry (scattering wavevector q perpendicular to the direction of flow) the measured relaxation times are independent of the flow rate and determined only by the diffusive motion of the particles. This is not generally valid and, in particular, for a `longitudinal flow' ( q ∥ flow) scattering geometry the relaxation times are strongly affected by the flow‐induced motion of the particles. The results here show that the Brownian diffusion of colloidal particles can be measured in a flowing sample and that, up to flux limitations, the experimental conditions under which this is possible are easier to achieve at higher values of q.     

X‐Treme beamline at SLS: X‐ray magnetic circular and linear dichroism at high field and low temperature

X‐Treme is a soft X‐ray beamline recently built in the Swiss Light Source at the Paul Scherrer Institut in collaboration with École Polytechnique Fédérale de Lausanne. The beamline is dedicated to polarization‐dependent X‐ray absorption spectroscopy at high magnetic fields and low temperature. The source is an elliptically polarizing undulator. The end‐station has a superconducting 7 T–2 T vector magnet, with sample temperature down to 2 K and is equipped with an in situ sample preparation system for surface science. The beamline commissioning measurements, which show a resolving power of 8000 and a maximum flux at the sample of 4.7 × 10¹² photons s^?1, are presented. Scientific examples showing X‐ray magnetic circular and X‐ray magnetic linear dichroism measurements are also presented.