Teaching about the birth of synchrotron light: the role of Frascati and a missed opportunity.

The users of synchrotron light are now tens of thousands throughout the world. Paradoxically, many of them do not know much about the early history of their domain. This is regrettable, since education about the initial developments makes it easier to fully understand synchrotron radiation and effectively use its amazing features. Scarcely known, in particular, is the key role of scientists working in Frascati, Italy. Partly based on his personal experiences, the author reports here relevant aspects of this story, including a pioneering French-Italian experiment that started in the early 1960s, and the Frascati contributions in the 1970s and 1980s to the birth of synchrotron light research. Finally, the unwise strategic decisions that prevented Italy from achieving absolute leadership in this domain - in spite of its unique initial advantages - are analyzed.

X-ray-induced Cu deposition and patterning on insulators at room temperature.

X-ray irradiation is shown to trigger the deposition of Cu from solution, at room temperature, on a wide variety of insulating substrates: glass, passivated Si, TiN/Ti/SiO2/Si and photoresists like PMMA and SU-8. The process is suitable for patterning and the products can be used as seeds for electroplating of thicker overlayers.

Ancient administrative handwritten documents: X-ray analysis and imaging.

Handwritten characters in administrative antique documents from three centuries have been detected using different synchrotron X-ray imaging techniques. Heavy elements in ancient inks, present even for everyday administrative manuscripts as shown by X-ray fluorescence spectra, produce attenuation contrast. In most cases the image quality is good enough for tomography reconstruction in view of future applications to virtual page-by-page `reading'. When attenuation is too low, differential phase contrast imaging can reveal the characters from refractive index effects. The results are potentially important for new information harvesting strategies, for example from the huge Archivio di Stato collection, objective of the Venice Time Machine project.

X-ray irradiation synthesis of PEG-coated Au-Pd nanoparticles.

We demonstrate that the combination of x-ray irradiation and capping by polyethylene glycol (PEG) produces excellent flexibility in controlling the structure of Au-Pd nanoparticles while preserving their catalytic performance. We specifically adopted two different fabrication methods: co-reduction and seed-assisted reduction. In both cases, precursor composition plays an important role in controlling the phases and size of the bimetallic nanoparticles. The optimal catalytic performance is obtained with the highest Pd concentration and when the nanoparticles consist of a Au core and a Pd shell.

Gold nanoparticles as multimodality imaging agents for brain gliomas.

BACKGROUND: Nanoparticles can be used for targeted drug delivery, in particular for brain cancer therapy. However, this requires a detailed analysis of nanoparticles from the associated microvasculature to the tumor, not easy because of the required high spatial resolution. The objective of this study is to demonstrate an experimental solution of this problem, based in vivo and post-mortem whole organ imaging plus nanoscale 3-dimensional (3D) X-ray microscopy. RESULTS: The use of gold nanoparticles (AuNPs) as contrast agents paved the way to a detailed high-resolution three dimensional (3D) X-ray and fluorescence imaging analysis of the relation between xenografted glioma cells and the tumor-induced angiogenic microvasculature. The images of the angiogenic microvessels revealed nanoparticle leakage. Complementary tests showed that after endocytotic internalization fluorescent AuNPs allow the visible-light detection of cells. CONCLUSIONS: AuNP-loading of cells could be extended from the case presented here to other imaging techniques. In our study, they enabled us to (1) identify primary glioma cells at inoculation sites in mice brains; (2) follow the subsequent development of gliomas. (3) Detect the full details of the tumor-related microvasculature; (4) Finding leakage of AuNPs from the tumor-related vasculature, in contrast to no leakage from normal vasculature.

Firefly light flashing: oxygen supply mechanism.

Firefly luminescence is an intriguing phenomenon with potential technological applications, whose biochemistry background was only recently established. The physics side of this phenomenon, however, was still unclear, specifically as far as the oxygen supply mechanism for light flashing is concerned. This uncertainty is due to the complex microscopic structure of the tracheal system: without fully knowing its geometry, one cannot reliably test the proposed mechanisms. We solved this problem using synchrotron phase contrast microtomography and transmission x-ray microscopy, finding that the oxygen consumption corresponding to mitochondria functions exceeds the maximum rate of oxygen diffusion from the tracheal system to the photocytes. Furthermore, the flashing mechanism uses a large portion of this maximum rate. Thus, the flashing control requires passivation of the mitochondria functions, e.g., by nitric oxide, and switching of the oxygen supply from them to photoluminescence.

X-ray microscopy and tomography detect the accumulation of bare and PEG-coated gold nanoparticles in normal and tumor mouse tissues.

We demonstrate that, with appropriate staining, high-resolution X-ray microscopy can image complicated tissue structures--cerebellum and liver--and resolve large or small amounts of Au nanoparticles in these tissues. Specifically, images of tumor tissue reveal high concentrations of accumulated Au nanoparticles. PEG (poly(ethylene glycol)) coating is quite effective in enhancing this accumulation and significantly modifies the mechanism of uptake by reticuloendothelial system (RES) organs.

A simplified description of X-ray free-electron lasers.

It is shown that an elementary semi-quantitative approach explains essential features of the X-ray free-electron laser mechanism, in particular those of the gain and saturation lengths. Using mathematical methods and derivations simpler than complete theories, this treatment reveals the basic physics that dominates the mechanism and makes it difficult to realise free-electron lasers for short wavelengths. This approach can be specifically useful for teachers at different levels and for colleagues interested in presenting X-ray free-electron lasers to non-specialized audiences.

Full-field microimaging with 8 keV X-rays achieves a spatial resolutions better than 20 nm.

Fresnel zone plates (450 nm thick Au, 25 nm outermost zone width) used as objective lenses in a full field transmission reached a spatial resolution better than 20 nm and 1.5% efficiency with 8 keV photons. Zernike phase contrast was also realized without compromising the resolution. These are very significant achievements in the rapid progress of high-aspect-ratio zone plate fabrication by combined electron beam lithography and electrodeposition.

Hard x-ray Zernike microscopy reaches 30 nm resolution.

Since its invention in 1930, Zernike phase contrast has been a pillar in optical microscopy and more recently in x-ray microscopy, in particular for low-absorption-contrast biological specimens. We experimentally demonstrate that hard-x-ray Zernike microscopy now reaches a lateral resolution below 30 nm while strongly enhancing the contrast, thus opening many new research opportunities in biomedicine and materials science.

One-pot tuning of Au nucleation and growth: from nanoclusters to nanoparticles.

We describe a simple and effective method to obtain colloidal surface-functionalized Au nanoparticles. The method is primarily based on irradiation of a gold solution with high-flux X-rays from a synchrotron source in the presence of 11-mercaptoundecanoic acid (MUA). Extensive tests of the products demonstrated high colloidal density as well as excellent stability, shelf life, and biocompatibility. Specific tests with X-ray diffraction, UV-visible spectrometry, visible microscopy, Fourier transform infrared spectroscopy, dark-field visible-light scattering microscopy, and transmission electron microscopy demonstrated that MUA, being an effective surfactant, not only allows tunable size control of the nanoparticles, but also facilitates functionalization. The nanoparticle sizes were 6.45 ± 1.58, 1.83 ± 1.21, 1.52 ± 0.37 and 1.18 ± 0.26 nm with no MUA and with MUA-to-Au ratios of 1:2, 1:1, and 3:1. The MUA additionally enabled functionalization with l-glycine. We thus demonstrated flexibility in controlling the nanoparticle size over a large range with narrow size distribution.

One-pot synthesis of AuPt alloyed nanoparticles by intense x-ray irradiation.

We synthesized AuPt alloyed nanoparticles in colloidal solution by a one-pot procedure based on synchrotron x-ray irradiation in the presence of PEG (polyethylene glycol). The exclusive presence of alloyed nanoparticles with fcc structure was confirmed by several different experiments including UV-vis spectroscopy, x-ray diffraction (XRD) and transmission electron microscopy (TEM). The composition of the AuPt alloyed nanoparticles can be varied in a continuous fashion by simply varying the feed ratios of Au and Pt precursors. The nanoparticles exhibited colloidal stability and biocompatibility, important for potential applications.

Imaging the cellular uptake of tiopronin-modified gold nanoparticles.

Well-dispersed gold nanoparticles (NP) coated with tiopronin were synthesized by X-ray irradiation without reducing agents. High-resolution transmission electron microscopy shows that the average core diameters of the NPs can be systematically controlled by adjusting the tiopronin to Au mole ratio in the reaction. Three methods were used to study the NP uptake by cells: quantitative measurements by inductively coupled plasma mass spectrometry, direct imaging with high lateral resolution transmission electron microscopy and transmission X-ray microscopy. The results confirmed that the NP internalization mostly occurred via endocytosis and concerned the cytoplasm. The particles, in spite of their small sizes, were not found to arrive inside the cell nuclei. The synthesis without reducing agents and solvents increased the biocompatibility as required for potential applications in analysis and biomedicine in general.

Detecting small lung tumors in mouse models by refractive-index microradiology.

Refractive-index (phase-contrast) radiology was able to detect lung tumors less than 1 mm in live mice. Significant micromorphology differences were observed in the microradiographs between normal, inflamed, and lung cancer tissues. This was made possible by the high phase contrast and by the fast image taking that reduces the motion blur. The detection of cancer and inflammation areas by phase contrast microradiology and microtomography was validated by bioluminescence and histopathological analysis. The smallest tumor detected is less than 1 mm(3) with accuracy better than 1 × 10(-3) mm(3). This level of performance is currently suitable for animal studies, while further developments are required for clinical application.

Tailored Au nanorods: optimizing functionality, controlling the aspect ratio and increasing biocompatibility.

Monodisperse gold nanorods with high aspect ratio were synthesized by x-ray irradiation. Irradiation was first used to stimulate the creation of seeds. Afterward, nanorod growth was stimulated either by chemical reduction or again by x-ray irradiation. In the last case, the entire process took place without reducing agents. The shape of the final products could be controlled by modulating the intensity of the x-ray irradiation during the seed synthesis. In turn, the nanorod aspect ratio determines the absorption wavelength of the nanorods that can thus be optimized for different applications. Likewise, the aspect ratio influences the uptake of the nanorods by HeLa cells.

Synchrotron microangiography studies of angiogenesis in mice with microemulsions and gold nanoparticles.

We present an effective solution for the problem of contrast enhancement in phase-contrast microangiography, with the specific objective of visualising small (<8 microm) vessels in tumor-related microangiogenesis. Different hydrophilic and hydrophobic contrast agents were explored in this context. We found that an emulsified version of the hydrophobic contrast agents Lipiodol provides the best contrast and minimal distortion of the circulation and vessel structure. Such emulsions are reasonably biocompatible and, with sizes of 0 +/- 0.8 microm, sufficient to diffuse to the smallest vessel and still provide reasonable contrast. We also explored the use of Au nanoparticle colloids that could be used not only to enhance contrast but also for interesting applications in nanomedicine. Both the Lipiodol microemulsions and Au nanoparticle colloids can be conjugated with medicines or cell specific labeling agents and their small size can allow the study of the diffusion of contrast agents through the vessel leakage. This enables direct imaging of drug delivery which is important for cancer treatment.

Simple dose rate measurements for a very high synchrotron X-ray flux.

Dose measurements based on methylene blue (MB) bleaching, widely used for ultraviolet light, can also be applied to X-rays including very high flux levels. This method has been tested by using both MB bleaching and Fricke dosimetry for a conventional monochromatic X-ray source and then for 'white-beam' synchrotron radiation. The results show that MB bleaching dosimetry can easily measure X-ray doses up to at least 10(5) Gy s(-1), as long as the MB concentration is sufficiently high. This condition can be verified from the deviations from linearity of the bleaching versus exposure time.

Solid-state photoelectron spectroscopy with synchrotron radiation.

Synchrotron radiation sources, providing intense, tunable, polarized, and stable beams of ultraviolet and x-ray photons, are having a great impact on biology, physics, chemistry, materials science, and other areas of research. Synchrotron radiation has revolutionized solid-state photoelectron spectroscopy by enhancing its capabilities for investigating the electronic behavior of solids and solid surfaces. Several fundamental photoemission techniques that rely on synchrotron radiation are discussed in this article. These techniques have an adjustable tunable surface sensitivity and provide the first direct mapping of the electronic band structure. Recent applications of photoelectron spectroscopy for studies of chemisorption geometries and surface structures are discussed.

Photoemission spectroscopy of the high-temperature superconductivity gap.

Superconductivity is related to the presence of a narrow forbidden gap in the spectrum of the possible energies for the electrons in the material. These "superconductivity gaps" have traditionally been studied with tunneling and infrared absorption experiments. A third, powerful technique has been made possible by the discovery of hightransition temperature materials: the direct observation of the gap in photoemission spectra. The data analysis requires a careful reconsideration of the standard Einstein-Fermi model of the photoelectric effect. The conclusions are surprisingly simple and offer an alternate way to measure superconductivity gaps. This approach can also be used to study the directional properties of the gap, phenomena related to the coherence length, and possible departures from Fermi-liquid behavior.

Temperature Dependence of the Superconducting Gap Anisotropy in Bi2Sr2CaCu2O8+x.

Detailed data on the momentum-resolved temperature dependence of the superconducting gap of Bi(2)Sr(2)CaCu(2)O(8+x) are presented, complemented by similar data on the intensity of the photoemission superconducting condensate spectral area. The gap anisotropy between the Gamma-Mand Gamma-X directions increases markedly with increasing temperature, contrary to what happens for conventional anisotropic-gap superconductors, such as lead. Specifically, the size of the superconducting gap along the Gamma-X direction decreases to values indistinguishable from zero at temperatures for which the gap retains virtually full value along the Gamma-M direction. These data rule out the simplest type of d-wave order parameter.