Quantitative micro-PIXE comparison of elemental distribution in Ni-hyperaccumulating and non-accumulating genotypes of Senecio coronatus

The Ni hyperaccumulator, plant species Senecio coronatus (Thunb.) Harv., Asteraceae is an example of plant adaptation mechanisms to different ecological conditions. This widespread species can inter alia be found on serpentine outcrops and the genotypes growing in serpentine soils show different ways of adaptation. The populations from two distant localities take up and translocate Ni in concentrations which are normally phytotoxic, while plants growing on a different site, in the vicinity of another hyperaccumulating species, absorb amounts which are typical for most of the plants found on serpentine soils. The NAC nuclear microprobe was used to compare the distribution of Ni and other elements in selected organs and cells with simultaneous use of PIXE and proton BackScattering (BS). Quantitative maps of stems showed large differences in concentrations and distributions of major and trace elements. In hyperaccumulating genotypes Ni is present everywhere within stem tissues, but the highest concentrations were found in the epidermis, cortex and phloem. In non-accumulating plants Ni was concentrated in the phloem. In the leaf epidermis Ni was concentrated in the cell walls for both accumulating and non-accumulating plants. These results suggest that biochemical diversity is more than morphological, because investigated genotypes belong to the same taxon.     

Accuracy of thick-target micro-PIXE analysis

The accuracy attainable in micro-PIXE analysis is assessed in terms of the X-ray production model and its assumptions, physical realities of the specimen, the necessary data base, and techniques of standardization. NTIS reference materials are analyzed to provide the experimental tests of accuracy.     

JAERI Takasaki in-air micro-PIXE system for various applications

In JAERI Takasaki, an in-air micro-PIXE system has been developed. This system enables multi-elemental mapping of samples in atmospheric environment with spatial resolution of 1 μm. Various research programs, such as biomedical research, dental study, environmental science and geology, have been started in recent years. Several related techniques for these applications were developed. A FTP server has been operated for the remote collaborators to share experimental data over the internet. We have developed image processing methods for elemental concentration analysis and a new beam monitoring technique in thick target irradiation.     

Lapis lazuli provenance study by means of micro-PIXE

In this paper we report about the micro-PIXE characterisation of lapis lazuli, for a provenance study of this semi-precious stone, used for glyptic as early as 7000 years ago. The final aim is to find markers permitting to identify the origin of the raw material coming from three quarries in regions of historical importance: Afghanistan, Pamir Mountains and Siberia. This may help to reconstruct trade routes, especially for ancient objects for which written testimonies are scanty or absent at all.Due to the heterogeneity of lapis lazuli we concentrate our attention on single phases instead of the whole stone; in particular we focused on two of the main phases: lazurite, responsible for the blue colour, and diopside, the most frequent accessory mineral. This study was preceded and completed by means of microanalysis with Scanning Electron Microscopy (SEM-EDX) and Cold-Cathodoluminescence (cold-CL) analysis.Despite the limited number of analysed samples, results are sufficient to exclude/suggest a few features as provenance markers, partly confirming what has been previously published in literature.     

Elemental micro-PIXE mapping of hypersensitive lesions in Lagenaria sphaerica (Cucurbitaceae) resistant to Sphaerotheca fuliginea (powdery mildew)

Genotypes of the Southern African cucurbit, Lagenaria sphaerica, that are resistant to powdery-mildew (Sphaerotheca fuliginea) exhibit foliar hypersensitive (HS) lesions on inoculation with this fungal pathogen. Elemental distributions across radially symmetrical HS lesions, surrounding unlesioned leaf tissue and uninoculated leaf tissue, were obtained using the true elemental imaging system (Dynamic Analysis) of the NAC Van de Graaff nuclear microprobe. Raster scans of 3 MeV protons were complemented by simultaneous PIXE and BS point analyses. The composition of cellulose (C₆H₁₀O₅) was used as constant matrix composition for scans, and the sample thickness was found from BS spectra. Si and elements heavier than Ca contributed to matrix composition within HS lesions and the locally elevated Ca raised the limits of detection for some trace metals of interest. In comparison to uninoculated tissue, inoculated tissue was characterised by higher overall concentrations of all measured elements except Cu. Fully developed, 6 day-old HS lesions and the surrounding tissue could be divided into five zones, centred on the fungal infection site. Each zone was characterized by distinct local elemental distributions (either depletion, or accumulation to potentially phytotoxic levels).     

Development of a micro-PIXE system using tapered glass capillary optics

The performance of a compact micro-PIXE system using a tapered glass capillary for beam focusing was examined with the help of Monte-Carlo simulations. The effective beam spot size was evaluated by an edge-scanning method and a two-dimensional element mapping was successfully demonstrated by using a copper fine-mesh target. The spatially resolved energy-spectrum measurement revealed that even protons scattered at the inner wall of the capillary have kinetic energies enough to induce the X-ray emission in the target. Monte-Carlo simulations showed that the beam-focusing factors of the capillaries used in this study are about 1.7. The uniformity of the beam does not depend so much on the distance from the capillary outlet, but the beam radius was found to be twice larger than its original size even at 1 mm from the capillary outlet, which was consistent with the effective beam spot size measurement.     

Red layered medieval stained glass window characterization by means of micro-PIXE technique

Red layered medieval stained glass windows on a transparent greenish substrate are characteristic of European medieval cathedrals, but few compositional analyses have been performed on the coloured layers. The PIXE technique has been performed on a red layered stained glass window obtained during the restoration works carried out in Las Huelgas Monastery in Burgos (Spain). Protons of 3 MeV with a beam of 4 × 5 μm² were used to acquire elemental maps of a cross section of the sample, in order to observe the homogeneity of the layered structure and its substrate.In our work, copper was detected as in other layered glasses but a correspondence with lower amounts of zinc has also been determined. Both elements appear enriched in the red coloured layers, while the other quantified elements have the same relative composition along the sample. Corrosion layers, due to the lead supporting structure of the window, were also found.     

Quantitative elemental localisation in leaves and stems of nickel hyperaccumulating shrub Hybanthusfloribundus subsp. floribundus using micro-PIXE spectroscopy

Hybanthusfloribundus (Lindl.) F.Muell. subsp. floribundus is a native Australian nickel (Ni) hyperaccumulating shrub and a promising species for rehabilitation and phytoremediation of Ni tailings. Spatial localisation and quantification of Ni in leaf and stem tissues of H.floribundus subsp. floribundus was studied using micro-proton-induced X-ray emission (micro-PIXE) spectroscopy. Young plants, grown in a potting mix under controlled glasshouse conditions were exposed to Ni concentrations of 0 and 26 mM kg⁻¹ for 20 weeks. Leaf and stem samples were hand-sectioned and freeze-dried prior to micro-PIXE analysis. Elemental distribution maps of leaves revealed Ni concentration of 7800 mg kg⁻¹ dry weight (DW) in whole leaf sections, which was identical to the bulk tissue analysis. Elemental maps showed that Ni was preferentially localised in the adaxial epidermis (10,000 mg kg⁻¹ DW) and reached a maximum of up to 10,000 mg kg⁻¹ DW in the leaf margin. Freeze-dried stem sections from the same plants contained lower Ni than leaf tissues (1800 mg kg⁻¹ versus 7800 mg kg⁻¹ DW, respectively), however did not resolve a clear pattern of compartmentalisation across different anatomical regions. Our results suggest localisation in epidermal cells is an important physiological mechanism involved in Ni accumulation and tolerance in leaves of H.floribundus subsp. floribundus.     

Elemental analysis of edible grains by micro-PIXE: Common buckwheat case study

The aim of this study was the adaptation of the micro-PIXE method for analysis of nutritionally relevant heavy elements in different tissues of the grain of common buckwheat (Fagopyrum esculentum), as a representative nutritionally interesting grain food source. At 57% of the buckwheat grain biomass, the endosperm was a modest nutrient source when compared to the cotyledons, at 17% of the biomass. These latter contained high concentrations of trace elements, representing 91% of the total grain Zn, 87% for P, 70% for S, 62% for Mg, 60% for K, 54% for Cu, 53% for Mn and 35% for Fe. The husk provided storage for 85% of the total Ca, 84% for Al, 83% for Si, 76% for Cl, 69% for Ti and 46% for Fe. Knowledge on these preferential elemental constitutions of the different grain tissues makes the possibility of designing target products with nutritionally optimal constitution more feasible. These data represent a basis for a more targeted approach to nutritional improvement of grains intended for human consumption.     

On the interpretation of micro-PIXE measurements on a prototype microstructured reference material

In order to determine the beam spot size and scanning properties of ion microbeam systems, a novel reference material has been developed, consisting of permalloy (81% Ni, 19% Fe) strip patterns on silicon substrate. Due to the choice of substrate and pattern materials, these samples exhibit a high elemental contrast suitable for analysis with X-ray detection and ion scattering techniques. The microlithographic production scheme is briefly described. A prototype chip of this material was investigated with PIXE and RBS analysis in a scanning nuclear microprobe. It proved to be extremely useful in the routine to focus the ion microbeam and to determine its spot size. Due to the microscopic structure of these samples, a geometric dependence of matrix effects in the production of Si X-rays from the substrate material could be shown. Even dead-time effects in the counting electronics, showing up as an apparent thickness gradient, could be observed. Besides its primary role in microbeam diagnostics, this reference material can serve an educational role in developing the analyst's ability to correctly identify and interpret such artefacts.     

External micro-PIXE analysis of Nd~(3+) accumulation in Euglena gracilis

External micro-PIXE measurements were done to investigate the accumulation of Nd~(3+)on green algae species euglena gracilis.According to the Nd distribution patterns in the gracilis cells,the biosorption of Nd~(3+)to the cell's compartments can be observed.Comparing elemental mappings of the cell treated with different doses of the 1 mg/mL Nd~(3+)solution,the Nd uptake of euglena gracilis cells do no relate with the doses.From distributions of Ca and Mg,it is found that the Ca is complementary to Nd partly,and the Nd and Mg distributions are alike to each other,showing that Nd may be mainly in the chlorophyll molecules.The biochemistry related is discussed.     

Micro-ERDA,micro-RBS and micro-PIXE techniques in the investigation of fish otoliths

Elemental distribution in the otolith of the fresh water fish burbot (Lota lota L.) collected in Hungary was measured with Elastic Recoil Detection Analysis (ERDA), Rutherford Backscattering Spectrometry (RBS) and as a complementary technique, Particle-Induced X-ray Emission (PIXE) with a focussed ion beam of 1.5 × 1.5 μm² spot size. The organic- and inorganic-rich regions of the otolith are distinguished and they are presented as hydrogen and calcium maps at depth regions of 0–70, 70–140 and 140–210 nm. The textured surface of the sample and its porosity were characterized from the effect on the RBS spectra. The oxygen and carbon PIXE elemental maps can also be used to identify the organic- and inorganic-rich regions of the otolith. The calcium map was found to be more homogeneous because the otolith structure is averaged in a larger depth. The trace elements Fe, Zn and Sr were detected only in very low concentration by micro-PIXE.     

Evaluation of specimen preparation techniques for micro-PIXE localisation of elements in hyperaccumulating plants

Hybanthus floribundus subsp. floribundus, a rare Australian Ni-hyperaccumulating shrub and Pityrogramma calomelanos var. austroamericana, an Australian naturalized As-hyperaccumulating fern are promising species for use in phytoremediation of contaminated sites. Micro-proton-induced X-ray emission (μ-PIXE) spectroscopy was used to map the elemental distribution of the accumulated metal(loid)s, Ca and K in leaf or pinnule tissues of the two plant species. Samples were prepared by two contrasting specimen preparation techniques: freeze-substitution in tetrahydrofuran (THF) and freeze-drying. The specimens were analysed to compare the suitability of each technique in preserving (i) the spatial elemental distribution and (ii) the tissue structure of the specimens. Further, the μ-PIXE results were compared with concentration of elements in the bulk tissue obtained by ICP-AES analysis.In H. floribundus subsp. floribundus, μ-PIXE analysis revealed Ni, Ca and K concentrations in freeze-dried leaf tissues were at par with bulk tissue concentrations. Elemental distribution maps illustrated that Ni was preferentially localised in the adaxial epidermal tissues (1% DW) and least concentration was found in spongy mesophyll tissues (0.53% DW). Conversely, elemental distribution maps of THF freeze-substituted tissues indicated significantly lower Ni, Ca and K concentrations than freeze-dried specimens and bulk tissue concentrations. Moreover, Ni concentrations were uniform across the whole specimen and no localisation was observed.In P. calomelanos var. austroamericana freeze-dried pinnule tissues, μ-PIXE revealed statistically similar As, Ca and K concentrations as compared to bulk tissue concentrations. Elemental distribution maps showed that As localisation was relatively uniform across the whole specimen. Once again, THF freeze-substituted tissues revealed a significant loss of As compared to freeze-dried specimens and the concentrations obtained by bulk tissue analysis.The results demonstrate that freeze-drying is a suitable sample preparation technique to study elemental distribution of ions in H. floribundus and P. calomelanos plant tissues using μ-PIXE spectroscopy. Furthermore, cellular structure was preserved in samples prepared using this technique.     

Investigation of impact materials around Barringer Meteor Crater by SEM-EDX and micro-PIXE techniques

Impact materials collected at the Barringer Meteor Crater have been characterized by SEM-EDX and micro-PIXE techniques. Fine textural and true elemental images were created. As a main feature silica-bearing shell and an S–Fe–Ni–Cu core could be distinguished. Three different types of S–Fe–Ni–Cu systems were identified such as chalcopyrite, pentlandite and pyrrhotite.     

Characterization of a millefiori glass find from Aquincum by SEM-EDX and micro-PIXE methods

Research has been focused on the analysis of archaeological glasses from the Roman age and medieval times. Study of ancient millefiori type glasses from the collection of Hungarian Museums has been started. A test measurement, carried out on a glass fragment supposedly part of a dish, was performed by SEM-EDX and micro-PIXE methods. Complementary analytical data were obtained for texture and composition. Results suggest that Roman and Mesopotamian techniques were used together. Our data contribute to data bases of millefiori glasses.     

Investigation of impact materials from the Barringer Meteor Crater by micro-XANES and micro-PIXE techniques

Impact materials from the Barringer Meteor Crater were examined by combined micro-X-ray absorption near edge structure (micro-XANES) and micro-particle induced X-ray emission (micro-PIXE) methods. Efforts were focussed on the complex characterization of their iron-rich inclusions. The lateral distribution of elements as well as the oxidation state of iron was determined. The study demonstrates the capabilities of chemical speciation screening based on energy selective micro-XRF maps in geology. With the help of this method zero-valent (metallic) and three-valent iron were excluded in the studied specimens without performing XANES in every pixel.     

Application of micro-PIXE to fish life history analyses: trace element analysis of otoliths

Otoliths are biogenic, carbonate concretions which form part of the hearing/balance system in fishes. The radial growth of otoliths and the variation of trace elements along the radius appear to capture important aspects of fishes' environmental history. At the Lund Nuclear Microprobe Laboratory, we have begun to use Proton-Induced X-ray Emission spectroscopy (PIXE) for micro-elemental analysis of otoliths. The experimental procedure is discussed and a number of examples of what can be investigated are presented. In particular, movement of diadromous species (eel, menhaden, and anadromous brown trout) can be detected between fresh and brackish water by Sr/Ca ratio. This technique has also been used to identify fish that were raised in freshwater hatcheries and then released to brackish water (pike-perch example).     

基于Micro-PIXE能谱的大气单颗粒物污染源模式识别研究

来自不同排放源的大气气溶胶,其化学组成特别是微量元素组成具有各自的特征,在Micro-PIXE上呈现不同的特征峰。因此单个气溶胶颗粒物的Micro-PIXE能谱特征可作为其指纹,查找污染源。本文提出了适应于Micro-PIXE谱的模式识别算法,并对这种以Micro-PIXE能谱特征溯源的方法进行了可靠性检验。结果表明以此种方法进行源解析具有较高的可信度。相应的模式识别算法具有准确,快速的特点。     

Three-dimensional measurement of elemental distribution in minute samples by combination of in-air micro-PIXE and STIM

A technique of measuring three-dimensional (3D) distribution of trace elements in a minute sample was studied using the in-air micro-PIXE (Particle Induced X-ray Emission) system. A 3D distribution was acquired by means of reconstructing numbers of projection images of the sample obtained by micro-PIXE analyses from different directions. This is basically the same technique as X-ray CT (Computed Tomography). However, when this technique was applied to the in-air micro-PIXE analysis, the X-ray yields should be corrected in taking account of such parameters as the X-ray production cross section, the X-ray attenuation coefficient, the energy of an incident particle, and the 3D densities of major elements in the analysis sample. In this study, STIM (Scanning Transmission Ion Microscopy)-CT was used to obtain the 3D density map of the sample. Corrected 3D distributions of sulfur and iron in a minute test sample were successfully measured with this technique in combination of STIM-CT and in-air micro-PIXE.     

Software masking for the accurate determination of elemental quantities in discrete areas of a micro-PIXE scan

A technique is described which allows X-ray energy spectra corresponding to discrete regions of a micro-PIXE scan recorded in multi-parameter mode to be generated. A software mask is created using both user-defined rectangular limits and an algorithm operating on the distribution of one of the major elements in the sample. This mask is used to select X-ray events originating in the region of interest, from which elemental concentrations can be calculated. This technique can allow accurate determination of element concentrations in irregular samples or regions of interest, enhance the signal-to-noise ratio for specimens on thick backings and also allows several areas of the sample to be analysed separately. The use of the technique is demonstrated in an experiment to quantify the total mass of barium contained in a single cell alga (Closterium tumidulum).