Makina Yabashi

High resolution-high energy x-ray photoelectron spectroscopy using third-generation synchrotron radiation source, and its application to Si-high k insulator systems

High-resolution x-ray photoelectron spectroscopy (XPS) at 6 keV photon energy has been realized utilizing high-flux-density x rays from the third generation high-energy synchrotron radiation facility, SPring-8. The method has been applied to analysis of high-k HfO2/interlayer/Si complementary metal–oxide–semiconductor gate-dielectric structures. With the high energy resolution and high throughput of our system, chemical-state differences were observed in the Si 1s, Hf 3d, and O 1s peaks for as-deposited and annealed samples. The results revealed that a SiOxNy interlayer is more effective in controlling the interface structure than SiO2. Our results show the wide applicability of high resolution XPS with hard x rays from a synchrotron source.

Efficient focusing of hard x rays to 25nm by a total reflection mirror

Nanofocused x rays are indispensable because they can provide high spatial resolution and high sensitivity for x-ray nanoscopy/spectroscopy. A focusing system using total reflection mirrors is one of the most promising methods for producing nanofocused x rays due to its high efficiency and energy-tunable focusing. The authors have developed a fabrication system for hard x-ray mirrors by developing elastic emission machining, microstitching interferometry, and relative angle determinable stitching interferometry. By using an ultraprecisely figured mirror, they realized hard x-ray line focusing with a beam width of 25nm at 15keV. The focusing test was performed at the 1-km-long beamline of SPring-8.

Determination of damage-free crystal structure of an X-ray–sensitive protein using an XFEL

We report a method of femtosecond crystallography for solving radiation damage–free crystal structures of large proteins at sub-angstrom spatial resolution, using a large single crystal and the femtosecond pulses of an X-ray free-electron laser (XFEL). We demonstrated the performance of the method by determining a 1.9-Å radiation damage–free structure of bovine cytochrome c oxidase, a large (420-kDa), highly radiation-sensitive membrane protein.

Beamline, experimental stations and photon beam diagnostics for the hard x-ray free electron laser of SACLA

A beamline for the x-ray free electron laser (XFEL) of SPring-8 Angstrom Compact free electron LAser (SACLA) provides hard x-ray pulses in the range 4.5–19.5 keV. Its optical system in an optics hutch delivers a pink beam below 15 keV with either of two double-mirror systems or a monochromatic beam with a double-crystal monochromator. These XFEL beams are used for various types of measurement at experimental stations, e.g. x-ray diffraction, coherent diffraction imaging, x-ray spectroscopy and pump-and-probe measurement. The experimental stations consist of experimental hutches and control stations, and a femtosecond optical laser which is synchronized with XFEL pulses. Photon diagnostics have been performed for measuring radiation parameters in a shot-by-shot manner.

Light-induced structural changes and the site of O=O bond formation in PSII caught by XFEL

Photosystem II (PSII) is a huge membrane-protein complex consisting of 20 different subunits with a total molecular mass of 350 kDa for a monomer. It catalyses light-driven water oxidation at its catalytic centre, the oxygen-evolving complex (OEC). The structure of PSII has been analysed at 1.9 Å resolution by synchrotron radiation X-rays, which revealed that the OEC is a Mn4CaO5 cluster organized in an asymmetric, ‘distorted-chair’ form. This structure was further analysed with femtosecond X-ray free electron lasers (XFEL), providing the ‘radiation damage-free’ structure. The mechanism of O=O bond formation, however, remains obscure owing to the lack of intermediate-state structures. Here we describe the structural changes in PSII induced by two-flash illumination at room temperature at a resolution of 2.35 Å using time-resolved serial femtosecond crystallography with an XFEL provided by the SPring-8 ångström compact free-electron laser. An isomorphous difference Fourier map between the two-flash and dark-adapted states revealed two areas of apparent changes: around the QB/non-haem iron and the Mn4CaO5 cluster. The changes around the QB/non-haem iron region reflected the electron and proton transfers induced by the two-flash illumination. In the region around the OEC, a water molecule located 3.5 Å from the Mn4CaO5 cluster disappeared from the map upon two-flash illumination. This reduced the distance between another water molecule and the oxygen atom O4, suggesting that proton transfer also occurred. Importantly, the two-flash-minus-dark isomorphous difference Fourier map showed an apparent positive peak around O5, a unique μ4-oxo-bridge located in the quasi-centre of Mn1 and Mn4 (refs 4,5). This suggests the insertion of a new oxygen atom (O6) close to O5, providing an O=O distance of 1.5 Å between these two oxygen atoms. This provides a mechanism for the O=O bond formation consistent with that proposed previously.

Development of an X-ray pixel detector with multi-port charge-coupled device for X-ray free-electron laser experiments

This paper presents development of an X-ray pixel detector with a multi-port charge-coupled device (MPCCD) for X-ray Free-Electron laser experiments. The fabrication process of the CCD was selected based on the X-ray radiation hardness against the estimated annual dose of 1.6 × 10(14) photon/mm(2). The sensor device was optimized by maximizing the full well capacity as high as 5 Me- within 50 μm square pixels while keeping the single photon detection capability for X-ray photons higher than 6 keV and a readout speed of 60 frames/s. The system development also included a detector system for the MPCCD sensor. This paper summarizes the performance, calibration methods, and operation status.

Grease matrix as a versatile carrier of proteins for serial crystallography

Serial femtosecond X-ray crystallography (SFX) has revolutionized atomic-resolution structural investigation by expanding applicability to micrometer-sized protein crystals, even at room temperature, and by enabling dynamics studies. However, reliable crystal-carrying media for SFX are lacking. Here we introduce a grease-matrix carrier for protein microcrystals and obtain the structures of lysozyme, glucose isomerase, thaumatin and fatty acid–binding protein type 3 under ambient conditions at a resolution of or finer than 2 Å.

Imaging live cell in micro-liquid enclosure by X-ray laser diffraction

Emerging X-ray free-electron lasers with femtosecond pulse duration enable single-shot snapshot imaging almost free from sample damage by outrunning major radiation damage processes. In bioimaging, it is essential to keep the sample close to its natural state. Conventional high-resolution imaging, however, suffers from severe radiation damage that hinders live cell imaging. Here we present a method for capturing snapshots of live cells kept in a micro-liquid enclosure array by X-ray laser diffraction. We place living Microbacterium lacticum cells in an enclosure array and successively expose each enclosure to a single X-ray laser pulse from the SPring-8 Angstrom Compact Free-Electron Laser. The enclosure itself works as a guard slit and allows us to record a coherent diffraction pattern from a weakly-scattering submicrometre-sized cell with a clear fringe extending up to a 28-nm full-period resolution. The reconstructed image reveals living whole-cell structures without any staining, which helps advance understanding of intracellular phenomena.

Microstitching interferometry for x-ray reflective optics

A new stitching interferometry based on a microscopic interferometer having peak-to-valley height accuracy of subnanometer order and lateral resolution higher than 20 μm was developed to measure surface figures of large-size x-ray mirror optics. Cumulative errors of the stitching angle in a long spatial wavelength range were effectively reduced to be 1×10−7 rad levels using another interferometer having a large cross section in the optical cavity. Some optical performances of ultraprecise x-ray mirrors, such as submicrofocused beam profile, were wave optically calculated from the measured surface figure profiles and observed at the 1 km long beamline (BL29XUL) of SPring-8. Observed and wave optically calculated results were in good agreement with a high degree of accuracy.

Relative angle determinable stitching interferometry for hard x-ray reflective optics

Metrology plays an important role in surface figuring with subnanometer accuracy. We have developed relative angle determinable stitching interferometry for the surface figuring of elliptical mirrors, in order to realize hard x-ray nanofocusing. In a stitching system, stitching angles are determined not by the general method using a common area between neighboring shots, but by the new method using the mirror’s tilt angles measured at times when profile data are acquired. The high measurement accuracy of approximately 4nm (peak-to-valley) was achieved in the measurement of a cylindrical surface having the same curvature as the elliptically designed shape to enable hard x-ray nanofocusing.Metrology plays an important role in surface figuring with subnanometer accuracy. We have developed relative angle determinable stitching interferometry for the surface figuring of elliptical mirrors, in order to realize hard x-ray nanofocusing. In a stitching system, stitching angles are determined not by the general method using a common area between neighboring shots, but by the new method using the mirror’s tilt angles measured at times when profile data are acquired. The high measurement accuracy of approximately 4nm (peak-to-valley) was achieved in the measurement of a cylindrical surface having the same curvature as the elliptically designed shape to enable hard x-ray nanofocusing.