| 1. |
- Bellucci, Valerio, et al.
(författare)
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Hard X-ray stereographic microscopy for single-shot differential phase imaging
- 2023
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Ingår i: Optics Express. - 1094-4087. ; 31:11, s. 18399-18406
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Tidskriftsartikel (refereegranskat)abstract
- The characterisation of fast phenomena at the microscopic scale is required for the understanding of catastrophic responses of materials to loads and shocks, the processing of materials by optical or mechanical means, the processes involved in many key technologies such as additive manufacturing and microfluidics, and the mixing of fuels in combustion. Such processes are usually stochastic in nature and occur within the opaque interior volumes of materials or samples, with complex dynamics that evolve in all three dimensions at speeds exceeding many meters per second. There is therefore a need for the ability to record three-dimensional X-ray movies of irreversible processes with resolutions of micrometers and frame rates of microseconds. Here we demonstrate a method to achieve this by recording a stereo phase-contrast image pair in a single exposure. The two images are combined computationally to reconstruct a 3D model of the object. The method is extendable to more than two simultaneous views. When combined with megahertz pulse trains of X-ray free-electron lasers (XFELs) it will be possible to create movies able to resolve 3D trajectories with velocities of kilometers per second.
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| 2. |
- Beyerlein, Kenneth R., et al.
(författare)
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Mix-and-diffuse serial synchrotron crystallography
- 2017
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Ingår i: IUCrJ. - : INT UNION CRYSTALLOGRAPHY. - 2052-2525. ; 4:6, s. 769-777
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Tidskriftsartikel (refereegranskat)abstract
- Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources.
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| 3. |
- Birnsteinova, Sarlota, et al.
(författare)
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Online dynamic flat-field correction for MHz microscopy data at European XFEL
- 2023
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Ingår i: Journal of Synchrotron Radiation. - 1600-5775. ; 30:6, s. 1030-1037
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Tidskriftsartikel (refereegranskat)abstract
- The high pulse intensity and repetition rate of the European X-ray Free-Electron Laser (EuXFEL) provide superior temporal resolution compared with other X-ray sources. In combination with MHz X-ray microscopy techniques, it offers a unique opportunity to achieve superior contrast and spatial resolution in applications demanding high temporal resolution. In both live visualization and offline data analysis for microscopy experiments, baseline normalization is essential for further processing steps such as phase retrieval and modal decomposition. In addition, access to normalized projections during data acquisition can play an important role in decision-making and improve the quality of the data. However, the stochastic nature of X-ray free-electron laser sources hinders the use of standard flat-field normalization methods during MHz X-ray microscopy experiments. Here, an online (i.e. near real-time) dynamic flat-field correction method based on principal component analysis of dynamically evolving flat-field images is presented. The method is used for the normalization of individual X-ray projections and has been implemented as a near real-time analysis tool at the Single Particles, Clusters, and Biomolecules and Serial Femtosecond Crystallography (SPB/SFX) instrument of EuXFEL.
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| 4. |
- Lieske, Julia, et al.
(författare)
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On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies
- 2019
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Ingår i: IUCrJ. - 2052-2525. ; 6:4, s. 714-728
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Tidskriftsartikel (refereegranskat)abstract
- Efficient and reliable sample delivery has remained one of the bottlenecks for serial crystallography experiments. Compared with other methods, fixed-target sample delivery offers the advantage of significantly reduced sample consumption and shorter data collection times owing to higher hit rates. Here, a new method of on-chip crystallization is reported which allows the efficient and reproducible growth of large numbers of protein crystals directly on micro-patterned silicon chips for in-situ serial crystallography experiments. Crystals are grown by sitting-drop vapor diffusion and previously established crystallization conditions can be directly applied. By reducing the number of crystal-handling steps, the method is particularly well suited for sensitive crystal systems. Excessive mother liquor can be efficiently removed from the crystals by blotting, and no sealing of the fixed-target sample holders is required to prevent the crystals from dehydrating. As a consequence, 'naked' crystals are obtained on the chip, resulting in very low background scattering levels and making the crystals highly accessible for external manipulation such as the application of ligand solutions. Serial diffraction experiments carried out at cryogenic temperatures at a synchrotron and at room temperature at an X-ray free-electron laser yielded high-quality X-ray structures of the human membrane protein aquaporin 2 and two new ligand-bound structures of thermolysin and the human kinase DRAK2. The results highlight the applicability of the method for future high-throughput on-chip screening of pharmaceutical compounds.
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| 5. |
- Reuter, Fabian, et al.
(författare)
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Laser-induced, single droplet fragmentation dynamics revealed through megahertz x-ray microscopy
- 2023
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Ingår i: Physics of Fluids. - 1070-6631. ; 35:11
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Tidskriftsartikel (refereegranskat)abstract
- The fragmentation dynamics of single water droplets from laser irradiation is studied with megahertz frame rate x-ray microscopy. Owed to the nearly refraction-free and penetrating imaging technique, we could look into the interior of the droplet and reveal that two mechanisms are responsible for the initial explosive fragmentation of the droplet. First, reflection and diffraction of the laser beam at the droplet interface result in the formation of laser ray caustics that lead to non-homogeneous heating of the droplet, locally above the critical temperature. Second, homogeneous cavitation in the droplet that is likely caused from shockwaves reflected as tension waves at the acoustic soft boundaries of the droplet. Further atomization occurs in three stages, first a fine sub-micrometer sized mist forms on the side of the droplet posterior to laser incidence, then micrometer sized droplets are expelled from the rim of an expanding liquid sheet, and finally into droplets of larger size through hole and ligament formation in the thinning liquid sheet where ligaments pinch off.
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| 6. |
- Vagovič, Patrik, et al.
(författare)
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Megahertz x-ray microscopy at x-ray free-electron laser and synchrotron sources
- 2019
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Ingår i: Optica. - 2334-2536. ; 6:9, s. 1106-1109
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Tidskriftsartikel (refereegranskat)abstract
- Modern emerging technologies, such as additive manufacturing, bioprinting, and new material production, require novel metrology tools to probe fundamental high-speed dynamics happening in such systems. Here we demonstrate the application of the megahertz (MHz) European X-ray Free-Electron Laser (EuXFEL) to image the fast stochastic processes induced by a laser on water-filled capillaries with micrometer-scale spatial resolution. The EuXFEL provides superior contrast and spatial resolution compared to equivalent state-of-the-art synchrotron experiments. This work opens up new possibilities for the characterization of MHz stochastic processes on the nanosecond to microsecond time scales with object velocities up to a few kilometers per second using XFEL sources.
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| 7. |
- Villanueva Perez, Pablo, et al.
(författare)
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Megahertz X-ray Multi-projection imaging
- 2023
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Ingår i: arXiv.org. - 2331-8422.
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- X-ray time-resolved tomography is one of the most popular X-raytechniques to probe dynamics in three dimensions (3D). Recent developments in time-resolved tomography opened the possibility of recordingkilohertz-rate 3D movies. However, tomography requires rotating thesample with respect to the X-ray beam, which prevents characterization of faster structural dynamics. Here, we present megahertz (MHz)X-ray multi-projection imaging (MHz-XMPI), a technique capable ofrecording volumetric information at MHz rates and micrometer resolution without scanning the sample. We achieved this by harnessing theunique megahertz pulse structure and intensity of the European X-rayFree-electron Laser with a combination of novel detection and reconstruction approaches that do not require sample rotations. Our approachenables generating multiple X-ray probes that simultaneously record several angular projections for each pulse in the megahertz pulse burst.We provide a proof-of-concept demonstration of the MHz-XMPI technique’s capability to probe 4D (3D+time) information on stochasticphenomena and non-reproducible processes three orders of magnitudefaster than state-of-the-art time-resolved X-ray tomography, by generating 3D movies of binary droplet collisions. We anticipate that MHz-XMPIwill enable in-situ and operando studies that were impossible before,either due to the lack of temporal resolution or because the systemswere opaque (such as for MHz imaging based on optical microscopy).
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