We present the first direct experimental confirmation of attosecond pulse generation in the hard X-ray regime with a free-electron laser. Our experiment is based on measurements of a nonlinear optical phenomenon known as amplified spontaneous emission (ASE) from 3d transition metals. By analyzing the yield of the collective X-ray fluorescence induced by ultrashort pulses at the Linac Coherent Light Source, we identify the generation of attosecond pulses and shot-to-shot fluctuations in their duration, ranging from 100 as to 400 as. The observed product of bandwidth and pulse duration for 100 as pulses is approximately 2 fs·eV, indicating the generation of nearly transform-limited pulses. Our results extend the photon energy reach of attosecond techniques by one order of magnitude, providing the ability to simultaneously probe matter on the time-scales of electronic phenomena and with atomic spatial resolution. Furthermore, attosecond hard X-ray pulses can outrun the fastest radiation damage processes, paving the way to single-shot damage-free X-ray measurements.

Attosecond hard x-ray pulses from the LCLS are focused to nanometer-sized spots using an advanced Kirkpatrick-Baez mirror system. Peak intensity at the focal point exceeds 1019 W/cm2, enabling a suite of damage-free nonlinear x-ray spectroscopy. (c) 2025 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement

The name of an author in the article by Walter et al. (2022) [J. Synchrotron Rad. 29, 957–968] is corrected.

The performance of metal and polymer foams used in inertial confinement fusion (ICF), inertial fusion energy (IFE), and high-energy-density (HED) experiments is currently limited by our understanding of their nanostructure and its variation in bulk material. We utilized an X-ray-free electron laser (XFEL) together with lensless X-ray imaging techniques to probe the 3D morphology of copper foams at nanoscale resolution (28 nm). The observed morphology of the thin shells is more varied than expected from previous characterizations, with a large number of them distorted, merged, or open, and a targeted mass density 14% less than calculated. This nanoscale information can be used to directly inform and improve foam modeling and fabrication methods to create a tailored material response for HED experiments.

With the nearly full spatial coherence of X-ray free electron lasers comes unprecedented requirements on the precise figure of X-ray mirrors. For example, the Time-resolved AMO Instrument (TMO) requires a bendable Kirkpatrick-Baez (KB) mirror system in order to produce a range of focus spot sizes, while maintaining nm-level figure error across the corresponding range of elliptical shapes. Here we describe the process of TMO KB mirror characterization using optical metrology, the detailed comparison with at-wavelength wavefront sensors during instrument commissioning, and the implementation of an automatic focusing system that ties together the wavefront sensor output with the mirror bender controls.

The goal of the dual KB mirror system for the CXI beamline of the LCLS-HE upgrade is to realize both micro-focusing and nano-focusing functions over a photon energy range of 7-21 keV. This system consists of two pairs of bendable mirrors with the length of 1 m. The dynamical range of the mirror bending is up to 104 due to the fine adjustment of the focusing length to 1 mm over a 10 m range. The prefigured shape and the width profile of each mirror are optimized to have good performance for both functions.

Cytochrome c oxidase (C c O) is a large membrane-bound hemeprotein that catalyzes the reduction of dioxygen to water. Unlike classical dioxygen binding hemeproteins with a heme b group in their active sites, C c O has a unique binuclear center (BNC) comprised of a copper atom (Cu B ) and a heme a 3 iron, where O 2 binds and is reduced to water. CO is a versatile O 2 surrogate in ligand binding and escape reactions. Previous time-resolved spectroscopic studies of the CO complexes of bovine C c O (bC c O) revealed that photolyzing CO from the heme a 3 iron leads to a metastable intermediate (Cu B -CO), where CO is bound to Cu B , before it escapes out of the BNC. Here, with a time-resolved serial femtosecond X-ray crystallography-based pump-probe method, we detected a geminate photoproduct of the bC c O-CO complex, where CO is dissociated from the heme a 3 iron and moved to a temporary binding site midway between the Cu B and the heme a 3 iron, while the locations of the two metal centers and the conformation of the Helix-X, housing the proximal histidine ligand of the heme a 3 iron, remain in the CO complex state. This new structure, combined with other reported structures of bC c O, allows the full definition of the ligand dissociation trajectory, as well as the associated protein dynamics.

The newly constructed time-resolved atomic, molecular and optical science instrument (TMO) is configured to take full advantage of both linear accelerators at SLAC National Accelerator Laboratory, the copper accelerator operating at a repetition rate of 120 Hz providing high per-pulse energy as well as the superconducting accelerator operating at a repetition rate of about 1 MHz providing high average intensity. Both accelerators power a soft X-ray free-electron laser with the new variable-gap undulator section. With this flexible light source, TMO supports many experimental techniques not previously available at LCLS and will have two X-ray beam focus spots in line. Thereby, TMO supports atomic, molecular and optical, strong-field and nonlinear science and will also host a designated new dynamic reaction microscope with a sub-micrometer X-ray focus spot. The flexible instrument design is optimized for studying ultrafast electronic and molecular phenomena and can take full advantage of the sub-femtosecond soft X-ray pulse generation program.

Multimeric protein assemblies are abundant in nature. Streptavidin is an attractive protein that provides a paradigm system to investigate the intra- and intermolecular interactions of multimeric protein complexes. Also, it offers a versatile tool for biotechnological applications. Here, we present two apo-streptavidin structures, the first one is an ambient temperature Serial Femtosecond X-ray crystal (Apo-SFX) structure at 1.7 Å resolution and the second one is a cryogenic crystal structure (Apo-Cryo) at 1.1 Å resolution. These structures are mostly in agreement with previous structural data. Combined with computational analysis, these structures provide invaluable information about structural dynamics of apo streptavidin. Collectively, these data further reveal a novel cooperative allostery of streptavidin which binds to substrate via water molecules that provide a polar interaction network and mimics the substrate biotin which displays one of the strongest affinities found in nature.

Many questions regarding dynamic materials could be answered by using time-resolved ultra-fast imaging techniques to characterize the physical and chemical behavior of materials in extreme conditions and their evolution on the nanosecond scale. In this work, we perform multi-frame phase-contrast imaging (PCI) of micro-voids in low density polymers under laser-driven shock compression. At the Matter in Extreme Conditions (MEC) Instrument at the Linac Coherent Light Source (LCLS), we used a train of four x-ray free electron laser (XFEL) pulses to probe the evolution of the samples. To visualize the void and shock wave interaction, we deployed the Icarus V2 detector to record up to four XFEL pulses, separated by 1-3 nanoseconds. In this work, we image elastic waves interacting with the micro-voids at a pressure of several GPa. Monitoring how the material’s heterogeneities, like micro-voids, dictate its response to a compressive wave is important for benchmarking the performances of inertial confinement fusion energy materials. For the first time in a single sample, we have combined an ultrafast x-ray framing camera and four XFEL pulse train to create an ultrafast movie of micro-void evolution under laser-driven shock compression. Eventually, we hope this technique will resolve the material density as it evolves dynamically under laser shock compression.

The COVID-19 pandemic has resulted in 198 million reported infections and more than 4 million deaths as of July 2021 (covid19.who.int). Research to identify effective therapies for COVID-19 includes: (1) designing a vaccine as future protection; (2) de novo drug discovery; and (3) identifying existing drugs to repurpose them as effective and immediate treatments. To assist in drug repurposing and design, we determine two apo structures of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease at ambient temperature by serial femtosecond X-ray crystallography. We employ detailed molecular simulations of selected known main protease inhibitors with the structures and compare binding modes and energies. The combined structural and molecular modeling studies not only reveal the dynamics of small molecules targeting the main protease but also provide invaluable opportunities for drug repurposing and structure-based drug design strategies against SARS-CoV-2.

Beam splitters and delay lines are among the key building blocks of modern-day optical laser technology. Progress in x-ray free electron laser source development and applications over the past decade is calling for their counterpart operating at the Angstrom wavelength regime. Recent efforts in x-ray optics development demonstrate relatively stable delay lines that most often adopt the division-of-wavefront approach for the beam splitting and recombination. However, the two exit beams in such configurations struggle to achieve sufficient mutual coherence to enable applications such as interferometry, correlation spectroscopy, and nonlinear spectroscopy. We present an experimental realization of the generation of highly mutually coherent pulse pairs using an amplitude-split delay line design based on transmission grating beam splitters and channel-cut crystals. The performance of the prototype system was analyzed in the context of x-ray coherent scattering and correlation spectroscopy, where nearly identical high-contrast speckle patterns from both branches were observed. We show in addition the high level of dynamical stability during continuous delay scans, a capability essential for high sensitivity ultrafast measurements.

For LCLS-II, we have developed fluorescence intensity monitors and power meters as intensity monitors. The Fluorescence Intensity Monitor (FIM) provides the non-invasive, pulse-by-pulse normalization of experiments. For the LCLS-II instruments, the diagnostic was constructed with an array of four microchannel plate assemblies and four avalanche photodiodes. The diagnostics are being installed in each Kirkpatrick Baez mirror chamber. The noise of the diagnostic will be evaluated against a goal of 1 %. The X-ray power meter delivers average power values. For the LCLS-II instruments, a power meter was selected compatible with high average power. In the LCLS-II instruments, power meters are being installed with each profile monitor in order to evaluate the transmission along the X-ray transport. A calibration of a set of power meters was carried out against a gas monitor detector at FLASH. In addition for all the power meters, a relative calibration was performed with a visible light source. At the endstations, a power meter will determined the pulse energy at the sample.

X-ray free-electron lasers (FELs) are being recognized as a powerful tool in an ever-increasing number of research fields, but are very limited as to the number of experiments that they can support. This work shows that more beamtime could be made available by using `parasitic' geometries, where a secondary experiment uses the X-ray beam that the primary experiment does not utilize. The first successful ptychography experiment, a scanning coherent diffractive imaging technique, in a parasitic geometry at an X-ray FEL is demonstrated. Utilizing the CXI hutch at the Linac Coherent Light Source (LCLS), it is shown that the obtained data are of high quality and that characterizing the beam using ptychography can be much faster than traditional imprinting methods.

The advancements in bright and coherent X-ray free-electron lasers (XFEL) have in the last decade revolutionized diverse fields of study, leading to an ever-increasing demand in more intense X-ray pulses. The accurate knowledge of the wavefront and the focal profile of such pulses ultimately dictates much of the experimental outcome. Here we present a single-shot shearing interferometry method to measure the wavefront of a focused hard XFEL pulse. Two identical transmission gratings are placed near the focus, and the interference pattern of overlapping diffraction orders is analyzed, resulting in a two-dimensional reconstruction of the X-ray wavefront. The spatial resolution and wavefront sensitivity can be tuned in-situ during the measurement. The method is non-invasive (i.e., the zeroth order or transmitted pulse is not modified in intensity or profile), allowing for either a simultaneous intensity measurement at high resolution or a fully characterized transmitted pulse for general experimental use.

The Macromolecular Femtosecond Crystallography (MFX) instrument at the Linac Coherent Light Source (LCLS) is the seventh and newest instrument at the world's first hard X-ray free-electron laser. It was designed with a primary focus on structural biology, employing the ultrafast pulses of X-rays from LCLSat atmospheric conditions to overcome radiation damage limitations in biological measurements. It is also capable of performing various time-resolved measurements. The MFX design consists of a versatile base system capable of supporting multiple methods, techniques and experimental endstations. The primary techniques supported are forward scattering and crystallography, with capabilities for various spectroscopic methods and time-resolved measurements. The location of the MFX instrument allows for utilization of multiplexing methods, increasing user access to LCLS by running multiple experiments simultaneously.

In this work we present the application of a 2D single grating wavefront sensor to align and characterize the 100 nm focus at the Coherent X-ray Imaging (CXI) endstation at the Linac Coherent Light Source (LCLS). The results agree well with a model of the system, indicating that the mirrors perform as designed when alignment is optimized. In addition, a comparison with the imprint technique confirms the validity of the results, which showed that wavefront-based alignment resulted in negligible astigmatism. Analysis of the retrieved focus profile indicates that intensities >10(21) W=cm(2) are achievable with currently available LCLS beam parameters and optimal mirror alignment.

The ribosome translates nucleotide sequences of messenger RNA to proteins through selection of cognate transfer RNA according to the genetic code. To date, structural studies of ribosomal decoding complexes yielding high-resolution data have predominantly relied on experiments performed at cryogenic temperatures. New lightsources like the X-ray free electron laser (XFEL) have enabled data collection from macromolecular crystals at ambient temperature. Here, we report an X-ray crystal structure of the Thermus thermophilus 30S ribosomal subunit decoding complex to 3.45 Å resolution using data obtained at ambient temperature at the Linac Coherent Light Source (LCLS). We find that this ambient-temperature structure is largely consistent with existing cryogenic-temperature crystal structures, with key residues of the decoding complex exhibiting similar conformations, including adenosine residues 1492 and 1493. Minor variations were observed, namely an alternate conformation of cytosine 1397 near the mRNA channel and the A-site. Our serial crystallography experiment illustrates the amenability of ribosomal microcrystals to routine structural studies at ambient temperature, thus overcoming a long-standing experimental limitation.