Single-exposure X-ray phase imaging microscopy with a grating interferometer.

The advent of hard X-ray free-electron lasers enables nanoscopic X-ray imaging with sub-picosecond temporal resolution. X-ray grating interferometry offers a phase-sensitive full-field imaging technique where the phase retrieval can be carried out from a single exposure alone. Thus, the method is attractive for imaging applications at X-ray free-electron lasers where intrinsic pulse-to-pulse fluctuations pose a major challenge. In this work, the single-exposure phase imaging capabilities of grating interferometry are characterized by an implementation at the I13-1 beamline of Diamond Light Source (Oxfordshire, UK). For comparison purposes, propagation-based phase contrast imaging was also performed at the same instrument. The characterization is carried out in terms of the quantitativeness and the contrast-to-noise ratio of the phase reconstructions as well as via the achievable spatial resolution. By using a statistical image reconstruction scheme, previous limitations of grating interferometry regarding the spatial resolution can be mitigated as well as the experimental applicability of the technique.

Maximum likelihood reconstruction for grating-based X-ray microscopy.

The combination of grating-based phase-contrast imaging with X-ray microscopy can result in a complicated image formation. Generally, transverse shifts of the interference fringes are nonlinearly dependent on phase differences of the measured wave front. We present an iterative reconstruction scheme based on a regularized maximum likelihood cost function that fully takes this dependency into account. The scheme is validated by numerical simulations. It is particularly advantageous at low photon numbers and when the premises for deconvolution-based reconstructions are not met. Our reconstruction scheme hence enables a broader applicability of X-ray grating interferometry in imaging and wave front sensing.

Familial chilblain lupus due to a gain-of-function mutation in STING.

OBJECTIVES: Familial chilblain lupus is a monogenic form of cutaneous lupus erythematosus caused by loss-of-function mutations in the nucleases TREX1 or SAMHD1. In a family without TREX1 or SAMHD1 mutation, we sought to determine the causative gene and the underlying disease pathology. METHODS: Exome sequencing was used for disease gene identification. Structural analysis was performed by homology modelling and docking simulations. Type I interferon (IFN) activation was assessed in cells transfected with STING cDNA using an IFN-ß reporter and Western blotting. IFN signatures in patient blood in response to tofacitinib treatment were measured by RT-PCR of IFN-stimulated genes. RESULTS: In a multigenerational family with five members affected with chilblain lupus, we identified a heterozygous mutation of STING, a signalling molecule in the cytosolic DNA sensing pathway. Structural and functional analyses indicate that mutant STING enhances homodimerisation in the absence of its ligand cGAMP resulting in constitutive type I IFN activation. Treatment of two affected family members with the Janus kinase (JAK) inhibitor tofacitinib led to a marked suppression of the IFN signature. CONCLUSIONS: A heterozygous gain-of-function mutation in STING can cause familial chilblain lupus. These findings expand the genetic spectrum of type I IFN-dependent disorders and suggest that JAK inhibition may be of therapeutic value.

Impacts of Underwater Noise on Marine Vertebrates: Project Introduction and First Results.

The project conducts application-oriented research on impacts of underwater noise on marine vertebrates in the North and Baltic Seas. In distinct subprojects, the hearing sensitivity of harbor porpoises and gray seals as well as the acoustic tolerance limit of harbor porpoises to impulsive noise from pile driving and stress reactions caused by anthropogenic noise is investigated. Animals are equipped with DTAGs capable of recording the actual surrounding noise field of free-swimming harbor porpoises and seals. Acoustic noise mapping including porpoise detectors in the Natura 2000 sites of the North and Baltic Seas will help to fully understand current noise impacts.

X-ray Dark-Field Imaging for Improved Contrast in Historical Handwritten Literature.

If ancient documents are too fragile to be opened, X-ray imaging can be used to recover the content non-destructively. As an extension to conventional attenuation imaging, dark-field imaging provides access to microscopic structural object information, which can be especially advantageous for materials with weak attenuation contrast, such as certain metal-free inks in paper. With cotton paper and different self-made inks based on authentic recipes, we produced test samples for attenuation and dark-field imaging at a metal-jet X-ray source. The resulting images show letters written in metal-free ink that were recovered via grating-based dark-field imaging. Without the need for synchrotron-like beam quality, these results set the ground for a mobile dark-field imaging setup that could be brought to a library for document scanning, avoiding long transport routes for valuable historic documents.

Noise Reduction for Single-Shot Grating-Based Phase-Contrast Imaging at an X-ray Backlighter.

X-ray backlighters allow the capture of sharp images of fast dynamic processes due to extremely short exposure times. Moiré imaging enables simultaneously measuring the absorption and differential phase-contrast (DPC) of these processes. Acquiring images with one single shot limits the X-ray photon flux, which can result in noisy images. Increasing the photon statistics by repeating the experiment to gain the same image is not possible if the investigated processes are dynamic and chaotic. Furthermore, to reconstruct the DPC and transmission image, an additional measurement captured in absence of the object is required. For these reference measurements, shot-to-shot fluctuations in X-ray spectra and a source position complicate the averaging of several reference images for noise reduction. Here, two approaches of processing multiple reference images in combination with one single object image are evaluated regarding the image quality. We found that with only five reference images, the contrast-to-noise ratio can be improved by approximately 13% in the DPC image. This promises improvements for short-exposure single-shot acquisitions of rapid processes, such as laser-produced plasma shock-waves in high-energy density experiments at backlighter X-ray sources such as the PHELIX high-power laser facility.

Three-dimensional imaging of xylem at cell wall level through near field nano holotomography.

Detailed imaging of the three-dimensionally complex architecture of xylary plants is important for studying biological and mechanical functions of woody plants. Apart from common two-dimensional microscopy, X-ray micro-computed tomography has been established as a three-dimensional (3D) imaging method for studying the hydraulic function of wooden plants. However, this X-ray imaging method can barely reach the resolution needed to see the minute structures (e.g. pit membrane). To complement the xylem structure with 3D views at the nanoscale level, X-ray near-field nano-holotomography (NFH) was applied to analyze the wood species Pinus sylvestris and Fagus sylvatica. The demanded small specimens required focused ion beam (FIB) application. The FIB milling, however, influenced the image quality through gallium implantation on the cell-wall surfaces. The measurements indicated that NFH is appropriate for imaging wood at nanometric resolution. With a 26 nm voxel pitch, the structure of the cell-wall surface in Pinus sylvestris could be visualized in genuine detail. In wood of Fagus sylvatica, the structure of a pit pair, including the pit membrane, between two neighboring fibrous cells could be traced tomographically.

Evaluation of the Weighted Mean X-ray Energy for an Imaging System Via Propagation-Based Phase-Contrast Imaging.

For imaging events of extremely short duration, like shock waves or explosions, it is necessary to be able to image the object with a single-shot exposure. A suitable setup is given by a laser-induced X-ray source such as the one that can be found at GSI (Helmholtzzentrum für Schwerionenforschung GmbH) in Darmstadt (Society for Heavy Ion Research), Germany. There, it is possible to direct a pulse from the high-energy laser Petawatt High Energy Laser for Heavy Ion eXperiments (PHELIX) on a tungsten wire to generate a picosecond polychromatic X-ray pulse, called backlighter. For grating-based single-shot phase-contrast imaging of shock waves or exploding wires, it is important to know the weighted mean energy of the X-ray spectrum for choosing a suitable setup. In propagation-based phase-contrast imaging the knowledge of the weighted mean energy is necessary to be able to reconstruct quantitative phase images of unknown objects. Hence, we developed a method to evaluate the weighted mean energy of the X-ray backlighter spectrum using propagation-based phase-contrast images. In a first step wave-field simulations are performed to verify the results. Furthermore, our evaluation is cross-checked with monochromatic synchrotron measurements with known energy at Diamond Light Source (DLS, Didcot, UK) for proof of concepts.

RPA and Rad51 constitute a cell intrinsic mechanism to protect the cytosol from self DNA.

Immune recognition of cytosolic DNA represents a central antiviral defence mechanism. Within the host, short single-stranded DNA (ssDNA) continuously arises during the repair of DNA damage induced by endogenous and environmental genotoxic stress. Here we show that short ssDNA traverses the nuclear membrane, but is drawn into the nucleus by binding to the DNA replication and repair factors RPA and Rad51. Knockdown of RPA and Rad51 enhances cytosolic leakage of ssDNA resulting in cGAS-dependent type I IFN activation. Mutations in the exonuclease TREX1 cause type I IFN-dependent autoinflammation and autoimmunity. We demonstrate that TREX1 is anchored within the outer nuclear membrane to ensure immediate degradation of ssDNA leaking into the cytosol. In TREX1-deficient fibroblasts, accumulating ssDNA causes exhaustion of RPA and Rad51 resulting in replication stress and activation of p53 and type I IFN. Thus, the ssDNA-binding capacity of RPA and Rad51 constitutes a cell intrinsic mechanism to protect the cytosol from self DNA.