Publications by authors named "Bajt S"

High-resolution X-ray imaging of noncrystalline objects is often achieved through the approach of scanning coherent diffractive imaging known as ptychography. The imaging resolution is usually limited by the scattering properties of the sample, where weak diffraction signals at the highest scattering angles compete with parasitic scattering. Here, we demonstrate that X-ray multilayer Laue lenses with a high numerical aperture (NA) can be used to create a strong reference beam that holographically boosts weak scattering from the sample over a large range of scattering angles, enabling high-resolution imaging that is tolerant of such background.

View Article and Find Full Text PDF

Multilayer Laue lenses are volume diffractive optical elements for hard X-rays with the potential to focus beams to sizes as small as 1 nm. This ability is limited by the precision of the manufacturing process, whereby systematic errors that arise during fabrication contribute to wavefront aberrations even after calibration of the deposition process based on wavefront metrology. Such aberrations can be compensated by using a phase plate.

View Article and Find Full Text PDF
Article Synopsis
  • The main protease (M) of SARS-CoV-2 is crucial for the virus's functionality and is considered a potential target for drug development, as it is only active in its reduced form.
  • When oxidized, M's activity halts but can be restored, indicating an evolutionary adaptation to oxidative environments, although the protective mechanisms haven't been fully elucidated.
  • Researchers determined the crystal structure of oxidized M, revealing a disulfide bond that affects its dimer stability and crystallization, providing insights into the protein's response to oxidative stress and its structural study conditions.*
View Article and Find Full Text PDF

Photolyase is an enzyme that uses light to catalyze DNA repair. To capture the reaction intermediates involved in the enzyme's catalytic cycle, we conducted a time-resolved crystallography experiment. We found that photolyase traps the excited state of the active cofactor, flavin adenine dinucleotide (FAD), in a highly bent geometry.

View Article and Find Full Text PDF

Free-electron lasers (FEL) are revolutionizing X-ray-based structural biology methods. While protein crystallography is already routinely performed at FELs, Small Angle X-ray Scattering (SAXS) studies of biological macromolecules are not as prevalent. SAXS allows the study of the shape and overall structure of proteins and nucleic acids in solution, in a quasi-native environment.

View Article and Find Full Text PDF

For decades, researchers have elucidated essential enzymatic functions on the atomic length scale by tracing atomic positions in real-time. Our work builds on possibilities unleashed by mix-and-inject serial crystallography (MISC) at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals.

View Article and Find Full Text PDF

The highest resolution of images of soft matter and biological materials is ultimately limited by modification of the structure, induced by the necessarily high energy of short-wavelength radiation. Imaging the inelastically scattered X-rays at a photon energy of 60 keV (0.02 nm wavelength) offers greater signal per energy transferred to the sample than coherent-scattering techniques such as phase-contrast microscopy and projection holography.

View Article and Find Full Text PDF

We demonstrate that x-ray fluorescence emission, which cannot maintain a stationary interference pattern, can be used to obtain images of structures by recording photon-photon correlations in the manner of the stellar intensity interferometry of Hanbury Brown and Twiss. This is achieved utilizing femtosecond-duration pulses of a hard x-ray free-electron laser to generate the emission in exposures comparable to the coherence time of the fluorescence. Iterative phasing of the photon correlation map generated a model-free real-space image of the structure of the emitters.

View Article and Find Full Text PDF

The results of an experimental study of micro-jets produced with a gas dynamic virtual nozzle (GDVN) under the influence of an electric field are provided and discussed for the first time. The experimental study is performed with a 50% volume mixture of water and ethanol, and nitrogen focusing gas. The liquid sample and gas Reynolds numbers range from 0.

View Article and Find Full Text PDF

For decades, researchers have been determined to elucidate essential enzymatic functions on the atomic lengths scale by tracing atomic positions in real time. Our work builds on new possibilities unleashed by mix-and-inject serial crystallography (MISC) at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals .

View Article and Find Full Text PDF

With the development of X-ray free-electron lasers (XFELs), producing pulses of femtosecond durations comparable with the coherence times of X-ray fluorescence, it has become possible to observe intensity-intensity correlations due to the interference of emission from independent atoms. This has been used to compare durations of X-ray pulses and to measure the size of a focusedX-ray beam, for example. Here it is shown that it is also possible to observe the interference of fluorescence photons through the measurement of the speckle contrast of angle-resolved fluorescence patterns.

View Article and Find Full Text PDF

In recent years, X-ray speckle tracking techniques have emerged as viable tools for wavefront metrology and sample imaging applications, and have been actively developed for use at synchrotron light sources. Speckle techniques can recover an image free of aberrations and can be used to measure wavefronts with a high angular sensitivity. Since they are compatible with low-coherence sources they can be also used with laboratory X-ray sources.

View Article and Find Full Text PDF
Article Synopsis
  • The European XFEL and LCLS II are powerful X-ray sources that can collect detailed data from crystals at rapid megahertz rates.
  • Researchers used these X-ray pulses to gather two complete datasets from a single lysozyme crystal in less than 1 microsecond, achieving high-resolution structures.
  • The comparison of these structures showed no radiation damage or significant changes, indicating that this multi-hit SFX technique can effectively capture fast structural changes in crystals.
View Article and Find Full Text PDF

Improvements in x-ray optics critically depend on the measurement of their optical performance. The knowledge of wavefront aberrations, for example, can be used to improve the fabrication of optical elements or to design phase correctors to compensate for these errors. At present, the characterization of such optics is made using intense x-ray sources, such as synchrotrons.

View Article and Find Full Text PDF

Serial femtosecond crystallography (SFX) is a powerful technique that exploits X-ray free-electron lasers to determine the structure of macro-molecules at room temperature. Despite the impressive exposition of structural details with this novel crystallographic approach, the methods currently available to introduce crystals into the path of the X-ray beam sometimes exhibit serious drawbacks. Samples requiring liquid injection of crystal slurries consume large quantities of crystals (at times up to a gram of protein per data set), may not be compatible with vacuum configurations on beamlines or provide a high background due to additional sheathing liquids present during the injection.

View Article and Find Full Text PDF

High-speed liquid micro-jets are used to rapidly and repeatedly deliver protein microcrystals to focused and pulsed X-ray beams in the method of serial femtosecond crystallography. However, the current continuous flow of crystals is mismatched to the arrival of X-ray pulses, wasting vast amounts of an often rare and precious sample. Here, we introduce a method to address this problem by periodically trapping and releasing crystals in the liquid flow, creating locally concentrated crystal bunches, using an optical trap integrated in the microfluidic supply line.

View Article and Find Full Text PDF
Article Synopsis
  • The study focuses on how enzymes interact with substrates or ligands in real time, specifically observing the first phase of the reaction using advanced imaging techniques.* -
  • Researchers utilized the European XFEL (EuXFEL) to achieve near-atomic resolution and track ceftriaxone binding to β-lactamase, combining high repetition rates and mix-and-inject technology for time-resolved measurements.* -
  • The findings included calculating a diffusion coefficient to understand concentrations in enzyme crystals over time and describing the binding of the inhibitor sulbactam, showcasing the potential of EuXFEL for biomedical research.*
View Article and Find Full Text PDF

Achromatic focusing systems for hard X-rays are examined which consist of a refractive lens paired with a diffractive lens. Compared with previous analyses, we take into account the behaviour of thick refractive lenses, such as compound refractive lenses and waveguide gradient index refractive lenses, in which both the focal length and the position of the principal planes vary with wavelength. Achromatic systems formed by the combination of such a thick refractive lens with a multilayer Laue lens are found that can operate at a focusing resolution of about 3 nm, over a relative bandwidth of about 1%.

View Article and Find Full Text PDF

The purpose of this work is to determine, based on the computational model, whether a mixture of a binary liquid is capable of producing longer, thinner and faster gas-focused micro-jets, compared to the mono-constituent liquids of its components. Mixtures of water with two different alcohols, water + ethanol and water + 2-propanol, are considered. The numerical study of pre-mixed liquids is performed in the double flow focusing nozzle geometry used in sample delivery in serial femtosecond crystallography experiments.

View Article and Find Full Text PDF

X-ray microscopy offers the opportunity to image biological and radiosensitive materials without special sample preparations, bridging optical and electron microscopy capabilities. However, the performance of such microscopes, when imaging radiosensitive samples, is not limited by their intrinsic resolution, but by the radiation damage induced on such samples. Here, we demonstrate a novel, to the best of our knowledge, radio-efficient microscope, scanning Compton X-ray microscopy (SCXM), which uses coherently and incoherently (Compton) scattered photons to minimize the deposited energy per unit of mass for a given imaging signal.

View Article and Find Full Text PDF

Liquid micro-jets are crucial for sample delivery of protein crystals and other macromolecular samples in serial femtosecond crystallography. When combined with MHz repetition rate sources, such as the European X-ray free-electron laser (EuXFEL) facility, it is important that the diffraction patterns are collected before the samples are damaged. This requires extremely thin and very fast jets.

View Article and Find Full Text PDF

Multilayer Laue lenses are diffractive optics for hard X-rays. To achieve high numerical aperture and resolution, diffracting structures of nanometer periods are required in such lenses, and a thickness (in the direction of propagation) of several micrometers is needed for high diffracting efficiency. Such structures must be oriented to satisfy Bragg's law, which can only be achieved consistently over the entire lens if the layers vary in their tilt relative to the incident beam.

View Article and Find Full Text PDF

Key insights in materials at extreme temperatures and pressures can be gained by accurate measurements that determine the electrical conductivity. Free-electron laser pulses can ionize and excite matter out of equilibrium on femtosecond time scales, modifying the electronic and ionic structures and enhancing electronic scattering properties. The transient evolution of the conductivity manifests the energy coupling from high temperature electrons to low temperature ions.

View Article and Find Full Text PDF

In recent years, X-ray speckle-tracking techniques have emerged as viable tools for wavefront metrology and sample imaging applications. These methods are based on the measurement of near-field images. Thanks to their simple experimental setup, high angular sensitivity and compatibility with low-coherence sources, these methods have been actively developed for use with synchrotron and laboratory light sources.

View Article and Find Full Text PDF
Article Synopsis
  • Serial femtosecond crystallography (SFX) using X-ray free electron lasers (XFELs) is a technique that helps in determining the structures of membrane proteins and observing changes over time, but traditional methods waste a lot of sample material.* -
  • The European XFEL produces rapid femtosecond X-ray pulses, but conventional liquid delivery methods result in over 99% sample wastage due to timing differences between pulse delivery.* -
  • A new microfluidic device that delivers protein crystal-laden droplets segmented by oil reduces sample waste by about 60%, allowing for the successful determination of a specific enzyme structure with previously unreported features.*
View Article and Find Full Text PDF