Observation of Histone H2AX Phosphorylation by Radiation-Induced Bystander Response Using Titanium Characteristic X-ray Microbeam.

Radiation-induced bystander response (RIBR) is a response induced in non-irradiated cells that receive bystander signals from directly irradiated cells. X-ray microbeams are useful tools for elucidating the mechanisms underlying RIBR. However, previous X-ray microbeams used low-energy soft X-rays with higher biological effects, such as aluminum characteristic X-rays, and the difference from conventional X-rays and γ-rays has often been discussed. The microbeam X-ray cell irradiation system at the Central Research Institute of Electric Power Industry has been upgraded to generate higher energy titanium characteristic X-rays (TiK X-rays), which have a longer penetration distance sufficient to irradiate 3D cultured tissues. Using this system, we irradiated the nuclei of HeLa cells with high precision and found that the pan-nuclear induction of phosphorylated histone H2AX on serine 139 (γ-H2AX) in the non-irradiated cells increased 180 and 360 min after irradiation. We established a new method to quantitatively evaluate bystander cells, using the fluorescence intensity of γ-H2AX as an indicator. The percentage of bystander cells increased significantly to 23.2% ± 3.2% and 29.3% ± 3.5% at 180 and 360 min after irradiation, respectively. Our irradiation system and the obtained results may be useful for studies of cell competition as well as non-targeted effects.

Performance of a ruthenium beam separator used to separate soft x rays from light generated by a high-order harmonic light source.

We describe the design and fabrication of a ruthenium beam separator used to simultaneously attenuate infrared light and reflect soft x rays. Measurements in the infrared and soft x-ray regions showed the beam separator to have a reflectivity of 50%-85% in the wavelength region from 6 to 10 nm at a grazing incidence angle of 7.5 deg and 4.3% at 800 nm and the same angle of grazing incidence, indicating that the amount of attenuation is 0.05-0.09. These results show that this beam separator could provide an effective means for separating IR light from soft x rays in light generated by high-order harmonic generation sources.

A proposal for a collecting mirror assembly for large divergence x-ray sources.

We propose a new type of collecting mirror assembly (CMA) for x rays, which will enable us to build a powerful optical system for collecting x rays from large divergence sources. The CMA consists of several mirror sections connected in series. The angle of each section is designed so that the x rays reflected from it are parallel to the x rays directly incident on the following sections. A simplified CMA structure is designed and applied to the Al-Kα emission line. It is estimated that by using the CMA the number of x rays detected could be increased by a factor of about 2.5.

Point spread function measurement of an X-ray beam focused by a multilayer zone plate with narrow annular aperture.

The experimental procedure for obtaining the point spread function (PSF) of a focusing beam generated using an X-ray multilayer zone plate (MZP) with a narrow annular aperture has been developed. It was possible to reconstruct the PSF by applying the tomographic process to the measured dataset consisting of line spread functions (LSFs) in every radial direction on the focal plane. The LSFs were measured by a knife-edge scanning method of detecting scattered intensity. In the experimental work, quasi-monochromatic undulator radiation with a first harmonic energy of 20 keV was directly focused without a monochromator by the MZP, and the PSF was measured using this procedure. As a result, a near diffraction-limited focused beam size of 46 nm full width at half-maximum was obtained.

Circular multilayer zone plate for high-energy x-ray nano-imaging.

A circular multilayer zone plate (MZP) was fabricated and its focusing performance was evaluated using 20-keV x-rays. MoSi(2) and Si layers were alternately deposited by DC magnetron sputtering on a wire core; all the interfaces satisfied the Fresnel zone condition. The measured line spread function was converted to a point spread function by tomographic reconstruction. The results suggest that the MZP has the potential to realize the diffraction-limited resolving power, which is calculated to be 35 nm using the diffraction integral. Furthermore, scanning transmission microscopy using the MZP could resolve a 50-nm line-and-space pattern.