Synchrotron‐radiation phase‐contrast imaging of human stomach and gastric cancer: in vitro studies

The electron density resolution of synchrotron‐radiation phase‐contrast imaging (SR‐PCI) is 1000 times higher than that of conventional X‐ray absorption imaging in light elements, through which high‐resolution X‐ray imaging of biological soft tissue can be achieved. For biological soft tissue, SR‐PCI can give better imaging contrast than conventional X‐ray absorption imaging. In this study, human resected stomach and gastric cancer were investigated using in‐line holography and diffraction enhanced imaging at beamline 4W1A of the Beijing Synchrotron Radiation Facility. It was possible to depict gastric pits, measuring 50–70 µm, gastric grooves and tiny blood vessels in the submucosa layer by SR‐PCI. The fine structure of a cancerous ulcer was displayed clearly on imaging the mucosa. The delamination of the gastric wall and infiltration of cancer in the submucosa layer were also demonstrated on cross‐sectional imaging. In conclusion, SR‐PCI can demonstrate the subtle structures of stomach and gastric cancer that cannot be detected by conventional X‐ray absorption imaging, which prompt the X‐ray diagnosis of gastric disease to the level of the gastric pit, and has the potential to provide new methods for the imageology of gastric cancer.     

Evaluation of the UFXC32k photon‐counting detector for pump–probe experiments using synchrotron radiation

This paper presents the performance of a single‐photon‐counting hybrid pixel X‐ray detector with synchrotron radiation. The camera was evaluated with respect to time‐resolved experiments, namely pump–probe–probe experiments held at SOLEIL. The UFXC camera shows very good energy resolution of around 1.5 keV and allows the minimum threshold setting to be as low as 3 keV keeping the high‐count‐rate capabilities. Measurements of a synchrotron characteristic filling mode prove the proper separation of an isolated bunch of photons and the usability of the detector in time‐resolved experiments.     

Non‐invasive measurement of X‐ray beam heating on a surrogate crystal sample

Cryocooling is a technique routinely used to mitigate the effects of secondary radiation damage on macromolecules during X‐ray data collection. Energy from the X‐ray beam absorbed by the sample raises the temperature of the sample. How large is the temperature increase and does this reduce the effectiveness of cryocooling? Sample heating by the X‐ray beam has been measured non‐invasively for the first time by means of thermal imaging. Specifically, the temperature rise of 1 mm and 2 mm glass spheres (sample surrogates) exposed to an intense synchrotron X‐ray beam and cooled in a laminar flow of nitrogen gas is experimentally measured. For the typical sample sizes, photon energies, fluxes, flux densities and exposure times used for macromolecular crystallographic data collection at third‐generation synchrotron radiation sources and with the sample accurately centered in the cryostream, the heating by the X‐ray beam is only a few degrees. This is not sufficient to raise the sample above the amorphous‐ice/crystalline‐ice transition temperature and, if the cryostream cools the sample to 100 K, not even enough to significantly enhance radiation damage from secondary effects.     

Morphological study of early‐stage lung cancer using synchrotron radiation

In the present study the feasibility of applying synchrotron radiation to the morphological study of early‐stage lung cancer has been investigated. Lewis lung cancer was implanted and grown in a nude mouse for different periods, and imaged using phase‐contrast synchrotron X‐rays. Morphological differences were clearly shown between the normal lung and cancerous tissues at this early stage. Irregular and tortuous angiogenesis were found in the periphery region of the developing lung cancer. Results from this study indicate that synchrotron X‐rays can be used for imaging cancer development and progression with minimal invasion.     

Tracking extended mucociliary transport activity of individual deposited particles: longitudinal synchrotron X‐ray imaging in live mice

To assess potential therapies for respiratory diseases in which mucociliary transit (MCT) is impaired, such as cystic fibrosis and primary ciliary dyskinesia, a novel and non‐invasive MCT quantification method has been developed in which the transit rate and behaviour of individual micrometre‐sized deposited particles are measured in live mice using synchrotron phase‐contrast X‐ray imaging. Particle clearance by MCT is known to be a two‐phase process that occurs over a period of minutes to days. Previous studies have assessed MCT in the fast‐clearance phase, ～20 min after marker particle dosing. The aim of this study was to non‐invasively image changes in particle presence and MCT during the slow‐clearance phase, and simultaneously determine whether repeat synchrotron X‐ray imaging of mice was feasible over periods of 3, 9 and 25 h. All mice tolerated the repeat imaging procedure with no adverse effects. Quantitative image analysis revealed that the particle MCT rate and the number of particles present in the airway both decreased with time. This study successfully demonstrated for the first time that longitudinal synchrotron X‐ray imaging studies are possible in live small animals, provided appropriate animal handling techniques are used and care is taken to reduce the delivered radiation dose.     

Focusing synchrotron radiation using a polycapillary half‐focusing X‐ray lens for imaging

An imaging system based on a polycapillary half‐focusing X‐ray lens (PHFXRL) and synchrotron radiation source has been designed. The focal spot size and the gain in power density of the PHFXRL were 22 µm (FWHM) and 4648, respectively, at 14.0 keV. The spatial resolution of this new imaging system was better than 5 µm when an X‐ray charge coupled device with a pixel size of 10.9 × 10.9 µm was used. A fossil of an ancient biological specimen was imaged using this system.     

Radiobiological features of the anti‐cancer strategies involving synchrotron X‐rays

Synchrotrons are opening new paths in innovative anti‐cancer radiotherapy strategies. Indeed, the fluence of X‐rays induced by synchrotrons is so high (10⁶ times higher than standard medical irradiators) that it enables the production of X‐ray beams tunable in energy (monochromatic beams) and in size (micrometric beams). Monochromatic synchrotron X‐ray beams theoretically permit photoactivate high‐Z elements to be introduced in or close to tumours in order to increase the yield of damage by enhanced energy photoabsorption. This is notably the case of attempts with iodinated contrast agents used in tumour imaging (the computed tomography therapy approach) and with platinated agents used in chemotherapy (the PAT‐Plat approach). Micrometric synchrotron X‐ray beams theoretically permit very high radiation doses to accumulate in tumours by using arrays of parallel microplanar beams that spare the surrounding tissues (the microbeam radiation therapy approach). These anti‐cancer applications of synchrotron radiation have been developed at the European Synchrotron Radiation Facility to be applied to glioma, one of the tumour tissues most refractory to standard treatments. In the present paper the molecular and cellular mechanisms involved in these three approaches are reviewed, in the context of recent advances in radiobiology. Furthermore, by considering the unavoidable biases, an attempt to propose a comparison of the different results obtained in preclinical trials dealing with rats bearing tumours is given.     

Radiation damage to protein specimens from electron beam imaging and diffraction: a mini‐review of anti‐damage approaches,with special reference to synchrotron X‐ray crystallography

Recent research progress using X‐ray cryo‐crystallography with the photon beams from third‐generation synchrotron sources has resulted in recognition that this intense radiation commonly damages protein samples even when they are held at 100 K. Other structural biologists examining thin protein crystals or single particle specimens encounter similar radiation damage problems during electron diffraction and imaging, but have developed some effective countermeasures. The aim of this concise review is to examine whether analogous approaches can be utilized to alleviate the X‐ray radiation damage problem in synchrotron macromolecular crystallography. The critical discussion of this question is preceded by presentation of background material on modern technical procedures with electron beam instruments using 300–400 kV accelerating voltage, low‐dose exposures for data recording, and protection of protein specimens by cryogenic cooling; these practical approaches to dealing with electron radiation damage currently permit best resolution levels of 6 Å (0.6 nm) for single particle specimens, and of 1.9 Å for two‐dimensional membrane protein crystals. Final determination of the potential effectiveness and practical value of using such new or unconventional ideas will necessitate showing, by experimental testing, that these produce significantly improved protection of three‐dimensional protein crystals during synchrotron X‐ray diffraction.     

Can soaked‐in scavengers protect metalloprotein active sites from reduction during data collection?

One of the first events taking place when a crystal of a metalloprotein is exposed to X‐ray radiation is photoreduction of the metal centres. The oxidation state of a metal cannot always be determined from routine X‐ray diffraction experiments alone, but it may have a crucial impact on the metal's environment and on the analysis of the structural data when considering the functional mechanism of a metalloenzyme. Here, UV–Vis microspectrophotometry is used to test the efficacy of selected scavengers in reducing the undesirable photoreduction of the iron and copper centres in myoglobin and azurin, respectively, and X‐ray crystallography to assess their capacity of mitigating global and specific radiation damage effects. UV–Vis absorption spectra of native crystals, as well as those soaked in 18 different radioprotectants, show dramatic metal reduction occurring in the first 60 s of irradiation with an X‐ray beam from a third‐generation synchrotron source. Among the tested radioprotectants only potassium hexacyanoferrate(III) seems to be capable of partially mitigating the rate of metal photoreduction at the concentrations used, but not to a sufficient extent that would allow a complete data set to be recorded from a fully oxidized crystal. On the other hand, analysis of the X‐ray crystallographic data confirms ascorbate as an efficient protecting agent against radiation damage, other than metal centre reduction, and suggests further testing of HEPES and 2,3‐dichloro‐1,4‐naphtoquinone as potential scavengers.     

Mercury in archeological hair samples from Xiongnu burials (Noin‐Ula,Mongolia): SR XRF and CXRM analysis

A technique has been developed for determining mercury content in the concentration range of 1–1000 μg/g in hair samples by X‐ray fluorescence analysis using synchrotron radiation (synchrotron radiation X‐ray fluorescence, Siberian Synchrotron and Terahertz Radiation Center, Budker Institute of Nuclear Physics SB RAS). The mercury content was identified in archeological hair samples from an ancient burial of Xiongnu nobility (Mongolia, mound 22, 1^st century BC–1^st century AD); the content values were elevated (up to 1100 μg/g) in all the samples (n = 41). An X‐ray microanalysis using polycapillary lenses in a confocal scheme (confocal X‐ray microscopy station) was developed at the Synchrotron radiation X‐ray fluorescence to establish mercury distribution in a cross section of hair shaft with a spatial resolution of 5 μm. The findings of the study make it possible to assume exogenous income of mercury (from the burial environment) to the hair.     

In situ removal of carbon contamination from optics in a vacuum ultraviolet and soft X‐ray undulator beamline using oxygen activated by zeroth‐order synchrotron radiation

Carbon contamination of optics is a serious issue in all soft X‐ray beamlines because it decreases the quality of experimental data, such as near‐edge X‐ray absorption fine structure, resonant photoemission and resonant soft X‐ray emission spectra in the carbon K‐edge region. Here an in situ method involving the use of oxygen activated by zeroth‐order synchrotron radiation was used to clean the optics in a vacuum ultraviolet and soft X‐ray undulator beamline, BL‐13A at the Photon Factory in Tsukuba, Japan. The carbon contamination of the optics was removed by exposing them to oxygen at a pressure of 10^?1–10^?4 Pa for 17–20 h and simultaneously irradiating them with zeroth‐order synchrotron radiation. After the cleaning, the decrease in the photon intensity in the carbon K‐edge region reduced to 2–5%. The base pressure of the beamline recovered to 10^?7–10^?8 Pa in one day without baking. The beamline can be used without additional commissioning.     

A multi‐MHz single‐shot data acquisition scheme with high dynamic range: pump–probe X‐ray experiments at synchrotrons

The technical implementation of a multi‐MHz data acquisition scheme for laser–X‐ray pump–probe experiments with pulse limited temporal resolution (100 ps) is presented. Such techniques are very attractive to benefit from the high‐repetition rates of X‐ray pulses delivered from advanced synchrotron radiation sources. Exploiting a synchronized 3.9 MHz laser excitation source, experiments in 60‐bunch mode (7.8 MHz) at beamline P01 of the PETRA III storage ring are performed. Hereby molecular systems in liquid solutions are excited by the pulsed laser source and the total X‐ray fluorescence yield (TFY) from the sample is recorded using silicon avalanche photodiode detectors (APDs). The subsequent digitizer card samples the APD signal traces in 0.5 ns steps with 12‐bit resolution. These traces are then processed to deliver an integrated value for each recorded single X‐ray pulse intensity and sorted into bins according to whether the laser excited the sample or not. For each subgroup the recorded single‐shot values are averaged over ～10⁷ pulses to deliver a mean TFY value with its standard error for each data point, e.g. at a given X‐ray probe energy. The sensitivity reaches down to the shot‐noise limit, and signal‐to‐noise ratios approaching 1000 are achievable in only a few seconds collection time per data point. The dynamic range covers 100 photons pulse^?1 and is only technically limited by the utilized APD.     

High‐efficiency coherence‐preserving harmonic rejection with crystal optics

This work reports a harmonic‐rejection scheme based on the combination of Si(111) monochromator and Si(220) harmonic‐rejection crystal optics. This approach is of importance to a wide range of X‐ray applications in all three major branches of modern X‐ray science (scattering, spectroscopy, imaging) based at major facilities, and especially relevant to the capabilities offered by the new diffraction‐limited storage rings. It was demonstrated both theoretically and experimentally that, when used with a synchrotron undulator source over a broad range of X‐ray energies of interest, the harmonic‐rejection crystals transmit the incident harmonic X‐rays on the order of 10^?6. Considering the flux ratio of fundamental and harmonic X‐rays in the incident beam, this scheme achieves a total flux ratio of harmonic radiation to fundamental radiation on the order of 10^?10. The spatial coherence of the undulator beam is preserved in the transmitted fundamental radiation while the harmonic radiation is suppressed, making this scheme suitable not only for current third‐generation synchrotron sources but also for the new diffraction‐limited storage rings where coherence preservation is an even higher priority. Compared with conventional harmonic‐rejection mirrors, where coherence is poorly preserved and harmonic rejection is less effective, this scheme has the added advantage of lower cost and footprint. This approach has been successfully utilized at the ultra‐small‐angle X‐ray scattering instrument at the Advanced Photon Source for scattering, imaging and coherent X‐ray photon correlation spectroscopy experiments. With minor modification, the harmonic rejection can be improved by a further five orders of magnitude, enabling even more performance capabilities.     

Hydrothermal formation of tobermorite studied by in situ X‐ray diffraction under autoclave condition

Hydrothermal formation of tobermorite from a pre‐cured cake has been investigated by transmission X‐ray diffraction (XRD) using high‐energy X‐rays from a synchrotron radiation source in combination with a newly designed autoclave cell. The autoclave cell has a large and thin beryllium window for wide‐angle X‐ray diffraction; nevertheless, it withstands a steam pressure of more than 1.2 MPa, which enables in situ XRD measurements in a temperature range of 373 to 463 K under a saturated steam pressure. Formation and/or decomposition of several components has been successfully observed during 7.5 h of reaction time. From the intensity changes of the intermediate materials, namely non‐crystalline C–S–H and hydroxylellestadite, two pathways for tobermorite formation have been confirmed. Thus, the newly developed autoclave cell can be used for the analyses of reaction mechanisms under specific atmospheres and temperatures.     

An in situ atomic force microscope for normal‐incidence nanofocus X‐ray experiments

A compact high‐speed X‐ray atomic force microscope has been developed for in situ use in normal‐incidence X‐ray experiments on synchrotron beamlines, allowing for simultaneous characterization of samples in direct space with nanometric lateral resolution while employing nanofocused X‐ray beams. In the present work the instrument is used to observe radiation damage effects produced by an intense X‐ray nanobeam on a semiconducting organic thin film. The formation of micrometric holes induced by the beam occurring on a timescale of seconds is characterized.     

Measurement of L subshell fluorescence yield ratios of some high Z elements by selective excitation method

The L₁, L₂ and L₃ subshells of Hf, Ta and Re atoms have been excited selectively by using microprobe XRF beam line, Indus‐2, RRCAT, India. The consequent characteristic L X‐ray photons, emitted from the targets due to creations of vacancies in L subshells, are measured using silicon drift detector (X‐123) spectrometer. As the energy of synchrotron radiation increases, the contribution of characteristic L X‐ray intensity increases. The advantage of the increase in the intensity of the characteristic L X‐ray photons with an increase in the energy of synchrotron radiation has been used to determine the L subshell fluorescence yield ratios of Hf, Ta and Re atoms by adopting the selective excitation method. The measured ratios of L subshell fluorescence yield have been compared with theoretical and other experimental values.     

Small‐angle pump–probe studies of photoexcited nanoparticles

X‐ray scattering experiments on femtosecond laser‐excited gold nanoparticle suspensions are presented. It is shown that the time‐resolved pump–probe technique using the X‐ray pulse structure at synchrotron sources is capable of resolving structural dynamics on the nanometer scale to high precision. The estimation of X‐ray flux density allows the projection of experiments on an X‐ray free‐electron laser probing single nanoparticles in a one‐shot exposure.     

The first microbeam synchrotron X‐ray fluorescence beamline at the Siam Photon Laboratory

The first microbeam synchrotron X‐ray fluorescence (µ‐SXRF) beamline using continuous synchrotron radiation from Siam Photon Source has been constructed and commissioned as of August 2011. Utilizing an X‐ray capillary half‐lens allows synchrotron radiation from a 1.4 T bending magnet of the 1.2 GeV electron storage ring to be focused from a few millimeters‐sized beam to a micrometer‐sized beam. This beamline was originally designed for deep X‐ray lithography (DXL) and was one of the first two operational beamlines at this facility. A modification has been carried out to the beamline in order to additionally enable µ‐SXRF and synchrotron X‐ray powder diffraction (SXPD). Modifications included the installation of a new chamber housing a Si(111) crystal to extract 8 keV synchrotron radiation from the white X‐ray beam (for SXPD), a fixed aperture and three gate valves. Two end‐stations incorporating optics and detectors for µ‐SXRF and SXPD have then been installed immediately upstream of the DXL station, with the three techniques sharing available beam time. The µ‐SXRF station utilizes a polycapillary half‐lens for X‐ray focusing. This optic focuses X‐ray white beam from 5 mm × 2 mm (H × V) at the entrance of the lens down to a diameter of 100 µm FWHM measured at a sample position 22 mm (lens focal point) downstream of the lens exit. The end‐station also incorporates an XYZ motorized sample holder with 25 mm travel per axis, a 5× ZEISS microscope objective with 5 mm × 5 mm field of view coupled to a CCD camera looking to the sample, and an AMPTEK single‐element Si (PIN) solid‐state detector for fluorescence detection. A graphic user interface data acquisition program using the LabVIEW platform has also been developed in‐house to generate a series of single‐column data which are compatible with available XRF data‐processing software. Finally, to test the performance of the µ‐SXRF beamline, an elemental surface profile has been obtained for a piece of ancient pottery from the Ban Chiang archaeological site, a UNESCO heritage site. It was found that the newly constructed µ‐SXRF technique was able to clearly distinguish the distribution of different elements on the specimen.     

Asynchronous sampling for ultrafast experiments with low momentum compaction at the ANKA ring

A high‐repetition‐rate pump–probe experiment is presented, based on the asynchronous sampling approach. The low‐α mode at the synchrotron ANKA can be used for a time resolution down to the picosecond limit for the time‐domain sampling of the coherent THz emission as well as for hard X‐ray pump–probe experiments, which probe structural dynamics in the condensed phase. It is shown that a synchronization of better than 1 ps is achieved, and examples of phonon dynamics of semiconductors are presented.     

Detection of circulating tumor cells via an X‐ray imaging technique

Detailed information on the location and the size of tumor cells circulating through lymphatic and blood vessels is useful to cancer diagnosis. Metastasis of cancers to other non‐adjacent organs is reported to cause 90% of deaths not from the primary tumors. Therefore, effective detection of circulating tumors cells (CTCs) related to metastasis is emphasized in cancer treatments. With the use of synchrotron X‐ray micro‐imaging techniques, high‐resolution images of individual flowing tumor cells were obtained. Positively charged gold nanoparticles (AuNPs) which were inappropriate for incorporation into human red blood cells were selectively incorporated into tumor cells to enhance the image contrast. This approach enables images of individual cancer cells and temporal movements of CTCs to be captured by the high X‐ray absorption efficiency of selectively incorporated AuNPs. This new technology for in vivo imaging of CTCs would contribute to improve cancer diagnosis and cancer therapy prognosis.