I18-Microfocus Spectroscopy
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Alexander P.
Morrell
,
Richard A.
Martin
,
Helen M
Roberts
,
Hiram
Castillo-Michel
,
J. Frederick W.
Mosselmans
,
Kalotina
Geraki
,
Adrian T.
Warfield
,
Paul
Lingor
,
Wasif
Qayyum
,
Daniel
Graf
,
Maria
Febbraio
,
Owen
Addison
Diamond Proposal Number(s):
[17638, 23569]
Open Access
Abstract: Exposures to exogenous particles is of increasing concern to human health. Characterising the concentrations, chemical species, distribution, and involvement of the stimulus with the tissue microanatomy is essential in understanding the associated biological response. However, no single imaging technique can interrogate all these features at once which confounds and limits correlative analyses. Developments of synchronous imaging strategies, allowing multiple features to be identified simultaneously, is essential to assess spatial relationships between these key features with greater confidence. Here we present data to first highlight complications of correlative analysis between the tissue microanatomy and elemental composition associated with imaging serial tissue sections. This is achieved by assessing both the cellular and elemental distribution in 3-dimensional space using optical microscopy on serial sections and confocal X-ray fluorescence spectroscopy on bulk samples respectively. We propose a new imaging strategy using lanthanide tagged antibodies with X-ray fluorescence spectroscopy. Using simulations, a series of lanthanide tags were identified as candidate labels for scenarios where tissue sections are imaged. The feasibility and value of the proposed approach is shown where an exposure of Ti was identified concurrently with CD45 positive cells at sub-cellular resolutions. Significant heterogeneity in the distribution of exogenous particles and cells can be present between immediately adjacent serial sections showing clear need of synchronous imaging methods. The proposed approach enables elemental compositions to be correlated with the tissue microanatomy in a highly multiplexed and non-destructive manner at high spatial resolutions with the opportunity for subsequent guided analysis.
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May 2023
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[17644]
Abstract: Analyses of the degraded mordant gilding on two early fourteenth-century paintings by Pietro Lorenzetti were undertaken at the UK synchrotron facility Diamond Light Source. Previous studies revealed a complex orpiment-tinted mordant and its application for adhering two separate layers of silver and gold leaf and demonstrated degradation of the original materials. A new study involving synchrotron radiation (SR) microfocus X-ray fluorescence (µ-XRF) maps on cross-sections of the discolored mordant allowed imaging of the extent of migration of mobile As-Ag-S bearing species and provided ideal positions from which to collect microfocus X-ray absorption near edge structure (µ-XANES) maps of As, Ag, and S for oxidation state analysis. SR µ-XANES indicates that arsenolite (As2O3) is present throughout the mordant suggesting light-induced photooxidation of orpiment. Both As3+ (as oxide) and As5+ have been detected and these are known signs for orpiment degradation. SR microfocus X-ray diffraction (µ-XRD) was used to reveal the nature of altered phases in the mordant layer and identified the presence of acanthite Ag2S, xanthoconite Ag3AsS3, and arsenolite As2O3. The analysis confirmed that the darkening of the mordant is most probably caused by finely dispersed grey-colored Ag + -bearing acanthite particles (Ag2S). The results are discussed in the context of analysis of contemporary paintings and recent studies on the alteration of orpiment.
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Apr 2023
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[23705]
Abstract: Background: Contrast agents (CA) are administered in magnetic resonance imaging (MRI) clinical exams to measure tissue perfusion, enhance image contrast between adjacent tissues, or provide additional biochemical information in molecular MRI. The efficacy of a CA is determined by the tissue distribution of the agent and its concentration in the extracellular space of all tissues. Methods: In this work, micro-synchrotron radiation x-ray fluorescence (µ-SRXRF) was used to examine and characterize a gadolinium-based zinc-sensitive agent (GdL2) currently under development for detection of prostate cancer (PCa) by MRI. Prostate tissue samples were collected from control mice and mice with known PCa after an MRI exam that included injection of GdL2. The samples were raster scanned to investigate trends in Zn, Gd, Cu, Fe, S, P, and Ca. Results: Significant Zn and Gd co-localization was observed in both healthy and malignant tissues. In addition, a marked decrease in Zn was found in the lateral lobe of the prostate obtained from mice with PCa. Conclusion: We demonstrate here that µ-SRXRF is a useful tool for monitoring the distribution of several elements including Zn and Gd in animal models of cancer. The optimized procedures for tissue preparation, processing, data collection, and analysis are described.
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Dec 2022
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[20436]
Open Access
Abstract: Diseases of the muscle tissue, particularly those disorders which result from the pathology of individual muscle cells, are often called myopathies. The diversity of the content of individual cells is of interest with regard to their role in both biochemical mechanisms and the structure of muscle tissue itself. These studies focus on the preliminary analysis of the differences that may occur between diseased tissues and tissues that have been recognised as a reference group. To do so, 13 samples of biopsied human muscle tissues were studied: 3 diagnosed as dystrophies, 6 as (non-dystrophic) myopathy and 4 regarded as references. From these sets of muscle biopsies, 135 completely measured muscle fibres were separated altogether, which were subjected to investigations using synchrotron radiation X-ray fluorescence (SR-XRF). Muscle fibres were analysed in terms of the composition of elements such as Br, Ca, Cl, Cr, Cu, Fe, K, Mn, P, S and Zn. The performed statistical tests indicate that all three groups (dystrophies—D; myopathies—M; references—R) show statistically significant differences in their elemental compositions, and the greatest impact, according to the multivariate discriminate analysis (MDA), comes from elements such as Ca, Cu, K, Cl and S.
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Jul 2022
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I18-Microfocus Spectroscopy
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Abstract: The leaching of heavy metals from post-industrial slag and other anthropogenic waste sites is detrimental for human health and the wider environment. Remediation of these sites can be costly and sustainable low carbon solutions are preferably sought. Examining natural analogues which stabilize metals could provide valuable insights into low-cost solutions to the legacy problems of aquatic environments that are impacted by leaching. Calcareous tufa, sometimes known as travertine limestone, forms naturally when calcium-rich groundwater is exchanged with atmospheric CO2 at mid to hyperalkaline pH resulting in a calcite (CaCO3) precipitation. Tufa has also been observed to form at a small number of old industrial sites (e.g. mining, steel works, paper mills) across northern England and Scotland. One site of interest is at Consett, N.E England, UK. Here tufa precipitates in the Howden Burn stream, a tributary of the River Derwent, as it emerges from the slag heaps from old steel work’s. Bulk analysis shows lead, arsenic, vanadium and zinc are present in the Howden Burn up to several 100 ppm. Analysis of the water downstream of the tufa shows metal concentrations are considerably reduced compared to concentrations upstream. High spatial resolution LA-ICP-MS analysis of the solid tufa sampled reveal metals present within the tufa structure. This leads to the hypothesis that the metals are precipitated together with the tufa during its formation. However, little is known about metal capture processes during tufa formation and the form that these metals are in. Here we present synchrotron micro X-ray fluorescence (μ-XRF) element maps of the tufa in cross-section that show the distributions of the metal within the laminations of the tufa structure. Understanding and exploitation of artificial tufa for metal capture could have potential as a CO2 positive solution for sustainable in-stream remediation.
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Mar 2022
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[24321]
Abstract: Looking for new green and environmentally friendly bio sorbents for metal removal from polluted wastewater, the present study investigates the potential new bio sorbent for Cd(II) removal from wastewater namely, the mechanism and uptake capacity of Cd(II) by brown algae, Fucus vesiculosus from the Irish Sea. This work takes a comprehensive approach involving the combination of qualitative and quantitative information collected from macro to atomistic scale, in a direct and non-destructive manner. Our results demonstrate that Cd(II) is adsorbed on the algal surface based on carboxylic of alginate groups. Effective Cd(II) adsorption is achieved at pH conditions between 5 and 7, at which the uptake occurs rapidly (∼2 h), with increasing Cd(II) concentration. Cd maximum uptake capacity (i.e., 1.203 mmol Cd g−1 dried algae) in first adsorption cycle show superior uptake as opposed to other species. Quantitatively the bio sorbent has an increasing uptake capacity (more than two folds) in the second cycle, after metal elution and biomass surface sites functioning. Desorption of Cd(II) and the regeneration of the biomass is effectively achieved with HCl (10 mM) and EDTA (1 mM), but they can only be used for two cycles, before the efficiency decreases. Microprecipitation occurs at high pH (>9) when using NaOH as an eluent. Results from this work shed new light on understanding Cd(II) binding mechanisms on Fucus v., providing crucial information for further process optimization, pilot testing, scaling up and implementation as a clean, environmentally friendly biotechnology applied to wastewater treatments.
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Oct 2021
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[20603]
Open Access
Abstract: Protein-based hybrid nanomaterials have recently emerged as promising platforms to fabricate tailored multifunctional biologics for biotechnological and biomedical applications. This work shows a simple, modular, and versatile strategy to design custom protein hybrid nanomaterials. This approach combines for the first time the engineering of a therapeutic protein module with the engineering of a nanomaterial-stabilizing module within the same molecule, resulting in a multifunctional hybrid nanocomposite unachievable through conventional material synthesis methodologies. As the first proof of concept, a multifunctional system was designed ad hoc for the therapeutic intervention and monitoring of myocardial fibrosis. This hybrid nanomaterial combines a designed Hsp90 inhibitory domain and a metal nanocluster stabilizing module resulting in a biologic drug labelled with a metal nanocluster. The engineered nanomaterial actively reduced myocardial fibrosis and heart hypertrophy in an animal model of cardiac remodeling. In addition to the therapeutic effect, the metal nanocluster allowed for in vitro, ex vivo, and in vivo detection and imaging of the fibrotic disease under study. This study evidences the potential of combining protein engineering and protein-directed nanomaterial engineering approaches to design custom nanomaterials as theranostic tools, opening up unexplored routes to date for the next generation of advanced nanomaterials in medicine.
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Dec 2020
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I18-Microfocus Spectroscopy
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Anouk M.
Borst
,
Martin P.
Smith
,
Adrian
Finch
,
Guillaume
Estrade
,
Cristina
Villanova-De-Benavent
,
Peter
Nason
,
Eva
Marquis
,
Nicola J.
Horsburgh
,
Kathryn M.
Goodenough
,
Cheng
Xu
,
Jindřich
Kynický
,
Kalotina
Geraki
Diamond Proposal Number(s):
[14793, 15903]
Open Access
Abstract: Global resources of heavy Rare Earth Elements (REE) are dominantly sourced from Chinese regolith-hosted ion-adsorption deposits in which the REE are inferred to be weakly adsorbed onto clay minerals. Similar deposits elsewhere might provide alternative supply for these high-tech metals, but the adsorption mechanisms remain unclear and the adsorbed state of REE to clays has never been demonstrated in situ. This study compares the mineralogy and speciation of REE in economic weathering profiles from China to prospective regoliths developed on peralkaline rocks from Madagascar. We use synchrotron X-ray absorption spectroscopy to study the distribution and local bonding environment of Y and Nd, as proxies for heavy and light REE, in the deposits. Our results show that REE are truly adsorbed as easily leachable 8- to 9-coordinated outer-sphere hydrated complexes, dominantly onto kaolinite. Hence, at the atomic level, the Malagasy clays are genuine mineralogical analogues to those currently exploited in China.
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Dec 2020
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I18-Microfocus Spectroscopy
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Diamond Proposal Number(s):
[7254]
Open Access
Abstract: In this work, the effects of the protozoan Neospora caninum on the bioenergetics, chemical composition, and elemental content of human brain microvascular endothelial cells (hBMECs) were investigated. We showed that N. caninum can impair cell mitochondrial (Mt) function and causes an arrest in host cell cycling at S and G2 phases. These adverse effects were also associated with altered expression of genes involved in Mt energy metabolism, suggesting Mt dysfunction caused by N. caninum infection. Fourier Transform Infrared (FTIR) spectroscopy analysis of hBMECs revealed alterations in the FTIR bands as a function of infection, where infected cells showed alterations in the absorption bands of lipid (2924 cm−1), amide I protein (1649 cm−1), amide II protein (1537 cm−1), nucleic acids and carbohydrates (1092 cm−1, 1047 cm−1, and 939 cm−1). By using quantitative synchrotron radiation X-ray fluorescence (μSR-XRF) imaging and quantification of the trace elements Zn, Cu and Fe, we detected an increase in the levels of Zn and Cu from 3 to 24 h post infection (hpi) in infected cells compared to control cells, but there were no changes in the level of Fe. We also used Affymetrix array technology to investigate the global alteration in gene expression of hBMECs and rat brain microvascular endothelial cells (rBMVECs) in response to N. caninum infection at 24 hpi. The result of transcriptome profiling identified differentially expressed genes involved mainly in immune response, lipid metabolism and apoptosis. These data further our understanding of the molecular events that shape the interaction between N. caninum and blood-brain-barrier endothelial cells.
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Aug 2020
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I18-Microfocus Spectroscopy
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Open Access
Abstract: Purpose : The purpose of this study was to investigate the distribution of Ca, Fe and Zn using X-ray fluorescence in human RPE/Bruch’s membrane/choroid with and without early AMD.
Methods : We used a set of unfixed frozen human retinal pigment epithelium (RPE)/choroid samples from young (n=1, female aged 34) and aged donors with (n=4, males, aged 73-78) and without (n=3, one female, aged 75-77) early AMD from Manchester Eye Tissue Repository. Using X-ray fluorescence microscopy (I18 Diamond light source, UK) with a 2 um beam, we obtained high-resolution Ca, Zn, Fe, sulphur, potassium, chloride and phosphorus maps covering areas up to 100 x 600 um2 of 30 um thick sections placed on quartz holders and scanned at room temperature
Results : Calcification was observed in the 3 groups. In the 34-year old sample, sparse small Ca spherules (2x2 um2) at the RPE/Bruch’s membrane interface not colocalising with Zn or Fe. In aged samples, with and without AMD, calcified nodules within RPE cells, at RPE/Bruch’s membrane interface and within druse in AMD. Every calcified nodule colocalised with Zn. Quantification revealed two types (high Zn content, low Zn content). In aged donors without AMD, high-Zn calcified nodules with Ca concentration of 3083+1679 ppm (mean+SD) (maximum 12809+9311) and Zn 66+55 ppm (max 125+75). The average size was 27+15 um2. In aged samples with AMD, high-Zn calcified nodules with average Ca concentration of 4316+1723 ppm (max 13105+7563) and Zn 97+57 ppm (max 201+143). Size 26x10 um2. In the aged non-AMD group, the low zinc-calcification nodules contained an average Ca ppm of 915+259 (max 1475+380) and Zn 33+18 ppm (max 40+21). The average size 28x16 um2. In the AMD group, the low Zn calcified nodules average Ca ppm 1544+1450 (max 3526+5289) and average Zn ppm 47+23 (max 64+37). Average size 19x10 um2. Calcified plaques in Bruch’s membrane from aged donors with and without AMD. Some of these plaques colocalised with Zn and also Fe. Fe-loaded structures in the choriocapillaris underlying calcified nodules and plaques.
Conclusions : Calcific nodules contain zinc in older eyes with and without AMD. Calcified nodules with lower amounts of Ca contained lower amounts of Zn, so the accumulation of Ca may occur in parallel to Zn. It is possible that iron-loaded structures in the choriocapillaris are macrophages.
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Jun 2020
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