Search Results (32)

Adipocytes derived from 3T3-L1 cells are a gold standard for analyses of adipogenesis processes and the metabolism of fat cells. A widely used histological and immunohistochemical staining and mass spectrometry lipidomics are mainly aimed for examining lipid droplets (LDs). Visualizing other cellular compartments contributing to the cellular machinery requires additional cell culturing for multiple labeling. Here, we present the localization of the intracellular structure of the 3T3-L1-derived adipocytes utilizing vibrational spectromicroscopy, which simultaneously illustrates the cellular compartments and provides chemical composition without extensive sample preparation and in the naïve state. Both vibrational spectra (FTIR—Fourier transform infrared and RS—Raman scattering spectroscopy) extended the gathered chemical information. We proved that both IR and RS spectra provide distinct chemical information about lipid content and their structure. Despite the expected presence of triacylglycerols and cholesteryl esters in lipid droplets, we also estimated the length and unsaturation degree of the fatty acid acyl chains that were congruent with known MS lipidomics of these cells. In addition, the clustering of spectral images revealed that the direct surroundings around LDs attributed to lipid-associated proteins and a high abundance of mitochondria. Finally, by using quantified markers of biomolecules, we showed that the fixative agents, paraformaldehyde and glutaraldehyde, affected the cellular compartment differently. We concluded that PFA preserves LDs better, while GA fixation is better for cytochromes and unsaturated lipid analysis. The proposed analysis of the spectral images constitutes a complementary tool for investigations into the structural and molecular features of fat cells. Full article

The acoustic communication system (ACS) in bark beetles has been studied mainly in species of the genera Dendroctonus, Ips and Polygraphus. Specifically, ACS of the roundheaded pine beetle, Dendroctonus adjunctus, has been little studied. In this study, we described the stridulatory apparatus of this beetle using optical and scanning electron microscopy and recorded the call types produced by males in three behavioral contexts: stress, female–male–, and male–male interactions. From the spectrograms and waveforms, call types, as well as temporal (tooth strike, tooth strike rate, and intertooth strike interval) and spectral features (minimum, maximum and dominant frequency), were determined. Males have a functional elytro–tergal stridulatory apparatus—females do not—consisting of a file for the pars stridens and two lobes for the plectrum. Most of spectro–temporal features were statistically different between single– and multi–noted calls and across the three behavioral contexts. In the male–male interaction, a new type of call named “withdrawal” was produced by the male withdrawing or fleeing. Our results suggest that the spectro–temporal features of single– and multiple–noted calls in the three behavioral conditions are specific and different from each other. Yet, the combination of single and multiple calls determines an overall calling pattern characteristic of the tested behaviors and, therefore, is species–specific. Full article

Modern catalysts are complex systems whose performance depends both on space and time domains and, most importantly, on the operational environment. As a direct consequence, understanding their functionalities requires sophisticated techniques and tools for measurement and simulation, addressing the proper spatial and temporal scale and being capable of mimicking the working conditions of every single component, such as catalyst supports, electrodes, electrolytes, as well as of the entire assembly, e.g., in the case of fuel cells or batteries. Scanning photoelectron spectro-microscopy (SPEM) is one of the approaches that allow combining X-ray photoelectron spectroscopy with sub-micron spatial resolution; in particular, the SPEM hosted at the ESCA Microscopy beamline at Elettra has been upgraded to conduct in situ and operando experiments. Three different case studies are presented to illustrate the capabilities of the SPEM in the investigation of catalytic materials in different conditions and processes. Full article

Changes in the structural properties of the skin due to collagen alterations are an important factor in diabetic skin complications. Using a combination of photonic methods as an optic diagnostic tool, we investigated the structural alteration in rat dermal collagen I in diabetes, and after short-term l-arginine treatment. The multiplex approach shows that in the early phase of diabetes, collagen fibers are partially damaged, resulting in the heterogeneity of fibers, e.g., “patchy patterns” of highly ordered/disordered fibers, while l-arginine treatment counteracts to some extent the conformational changes in collagen-induced by diabetes and mitigates the damage. Raman spectroscopy shows intense collagen conformational changes via amides I and II in diabetes, suggesting that diabetes-induced structural changes in collagen originate predominantly from individual collagen molecules rather than supramolecular structures. There is a clear increase in the amounts of newly synthesized proline and hydroxyproline after treatment with l-arginine, reflecting the changed collagen content. This suggests that it might be useful for treating and stopping collagen damage early on in diabetic skin. Our results demonstrate that l-arginine attenuates the early collagen I alteration caused by diabetes and that it could be used to treat and prevent collagen damage in diabetic skin at a very early stage. Full article

Highly aligned multi-wall carbon nanotubes were investigated with scanning electron microscopy (SEM), Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) before and after bombardment performed using noble gas ions of different masses (argon, neon and helium), in an ultra-high-vacuum (UHV) environment. Ion irradiation leads to change in morphology, deformation of the carbon (C) honeycomb lattice and different structural defects in multi-wall carbon nanotubes. One of the major effects is the production of bond distortions, as determined by micro-Raman and micro-X-ray photoelectron spectroscopy. We observe an increase in sp${}^3$ distorted bonds at higher binding energy with respect to the expected sp${}^2$ associated signal of the carbon 1s core level, and increase in dangling bonds. Furthermore, the surface damage as determined by the X-ray photoelectron spectroscopy carbon 1s core level is equivalent upon bombarding with ions of different masses, while the impact and density of defects in the lattice of the MWCNTs as determined by micro-Raman are dependent on the bombarding ion mass; heavier for helium ions, lighter for argon ions. These results on the controlled increase in sp${}^3$ distorted bonds, as created on the multi-wall carbon nanotubes, open new functionalization prospects to improve and increase atomic hydrogen uptake on ion-bombarded multi-wall carbon nanotubes. Full article

Thiomicrorhabdus species, belonging to the family Piscirickettsiaceae in the phylum Pseudomonadotav are usually detected in various sulfur-rich marine environments. However, only a few bacteria of Thiomicrorhabdus have been isolated, and their ecological roles and environmental adaptations still require further understanding. Here, we report a novel strain, XGS-01^T, isolated from a coastal sediment, which belongs to genus Thiomicrorhabdus and is most closely related to Thiomicrorhabdus hydrogeniphila MAS2^T, with a sequence similarity of 97.8%. Phenotypic characterization showed that XGS-01^T is a mesophilic, sulfur-oxidizing, obligate chemolithoautotrophy, with carbon dioxide as its sole carbon source and oxygen as its sole electron acceptor. During thiosulfate oxidation, strain XGS-01^T can produce extracellular sulfur of elemental α-S₈, as confirmed via scanning electron microscopy and Raman spectromicroscopy. Polyphasic taxonomy results indicate that strain XGS-01^T represents a novel species of the genus Thiomicrorhabdus, named Thiomicrorhabdus lithotrophica sp. nov. Genomic analysis confirmed that XGS-01^T performed thiosulfate oxidation through a sox multienzyme complex, and harbored fcc and sqr genes for sulfide oxidation. Comparative genomics analysis among five available genomes from Thiomicrorhabdus species revealed that carbon fixation via the oxidation of reduced-sulfur compounds coupled with oxygen reduction is conserved metabolic pathways among members of genus Thiomicrorhabdus. Full article

Solid-state batteries (SSB), characterized by solid-state electrolytes—in particular inorganic ones (ISSE)—are an ideal option for the safe implementation of metallic Li anodes. Even though SSBs with ISSEs have been extensively investigated over the last two decades, they still exhibit a series of technological drawbacks. In fact, mechano-chemical issues, mainly the stability of the electrolyte/anode interface, hinder their widespread application. The present investigation focusses on a thin-film LMO (Lithium-Manganese-Oxide)/LAGP (LiAlGe Phosphate)/Copper, anodeless Lithium-metal battery and explores the morphochemical evolution of the electrode/electrolyte interfaces with synchrotron-based Scanning Photoelectron Microscopy (SPEM) of intact pristine and cycled cells. Chemical images were acquired with submicrometer resolution, to highlight the coupled geometrical and chemical-state changes caused by electrochemical ageing. Geometrical changes of the electrolyte/cathode interface were induced by periodic volume changes, causing de-cohesion of the solid-solid contact, but no chemical-state changes accompany the cathodic damaging mode. Instead, shape changes of the electrolyte/anode region pinpoint the correlation between mechanical damaging with the decomposition of the LAGP ISSE, due to the reduction of Ge, triggered by the contact with elemental Li. The micro-spectroscopic approach adopted in this study enabled the assessment of the highly localized nature of the cathodic and anodic degradation modes in SSB devices and to single out the chemical and mechanical contributions. Full article

Micro electro-mechanical systems (MEMS) combining sensing and microfluidics functionalities, as are common in Lab-on-Chip (LoC) devices, are increasingly based on polymers. Benefits of polymers include tunable material properties, the possibility of surface functionalization, compatibility with many micro and nano patterning techniques, and optical transparency. Often, additional materials, such as metals, ceramics, or silicon, are needed for functional or auxiliary purposes, e.g., as electrodes. Hybrid patterning and integration of material composites require an increasing range of fabrication approaches, which must often be newly developed or at least adapted and optimized. Here, a microfabrication process concept is developed that allows one to implement attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and electrochemistry on an LoC device. It is designed to spatially resolve chemical sensitivity and selectivity, which are instrumental for the detection of chemical distributions, e.g., during on-flow chemical and biological reaction chemistry. The processing sequence involves (i) direct-write and soft-contact UV lithography in SUEX dry resist and replication in polydimethylsiloxane (PDMS) elastomers as the fluidic structure; (ii) surface functionalization of PDMS with oxygen plasma, 3-aminopropyl-triethoxysilane (APTES), and a UV-curable glue (NOA 73) for bonding the fluidic structure to the substrate; (iii) double-sided patterning of silicon nitride-coated silicon wafers serving as the ATR-FTIR-active internal reflection element (IRE) on one side and the electrode-covered substrate for microfluidics on the back side with lift-off and sputter-based patterning of gold electrodes; and (iv) a custom-designed active vacuum positioning and alignment setup. Fluidic channels of 100 μm height and 600 μm width in 5 mm thick PDMS were fabricated on 2” and 4” demonstrators. Electrochemistry on-chip functionality was demonstrated by cyclic voltammetry (CV) of redox reactions involving iron cyanides in different oxidation states. Further, ATR-FTIR measurements of laminar co-flows of H₂O and D₂O demonstrated the chemical mapping capabilities of the modular fabrication concept of the LoC devices. Full article

An online automatic feedback system has been successfully installed and commissioned at the BL08U1A Soft X-ray Spectromicroscopy Beamline of Shanghai Synchrotron Radiation Facility, which can monitor the incident X-ray beam in real time by measuring the blade-edge signals of the exit slit and automatically adjust the elliptical cylindrical mirror parameters to achieve beam calibration and maintain the optimal X-ray flux of the sample. This work provides a comprehensive description of the hardware composition, system implementation, feedback logic, function and software design, system optimization and commission, as well as the online experimental results supported by the system. The experimental results demonstrated that the online automatic feedback system is capable of effectively maintaining the optimal X-ray beam flux for X-ray absorption spectroscopy experiments. Its success can provide valuable technique assistance for the design, construction and optimization of similar systems at various beamlines in synchrotron sources in the future. Full article

Scanning transmission X-ray microscopy (STXM) provides the imaging of biological specimens allowing the parallel collection of localized spectroscopic information by X-ray fluorescence (XRF) and/or X-ray Absorption Near Edge Spectroscopy (XANES). The complex metabolic mechanisms which can take place in biological systems can be explored by these techniques by tracing even small quantities of the chemical elements involved in the metabolic pathways. Here, we present a review of the most recent publications in the synchrotrons’ scenario where soft X-ray spectro-microscopy has been employed in life science as well as in environmental research. Full article

In this report, we demonstrated the formation of a biomimetic mineralizing layer obtained on the surface of dental enamel (biotemplate) using bioinspired nanocrystalline carbonate-substituted calcium hydroxyapatite (ncHAp), whose physical and chemical properties are closest to the natural apatite dental matrix, together with a complex of polyfunctional organic and polar amino acids. Using a set of structural, spectroscopy, and advanced microscopy techniques, we confirmed the formation of a nanosized ncHAp-based mineralized layer, as well as studying its chemical, substructural, and morphological features by means of various methods for the pretreatment of dental enamel. The pretreatment of a biotemplate in an alkaline solution of Ca(OH)₂ and an amino acid booster, together with the executed subsequent mineralization with ncHAp, led to the formation of a mineralized layer with homogeneous micromorphology and the preferential orientation of the ncHAp nanocrystals. It was shown that the homogeneous crystallization of hydroxyapatite on the biotemplate surface and binding of individual nanocrystals and agglomerates into a single complex by an amino acid booster resulted in an increase (~15%) in the nanohardness value in the enamel rods area, compared to that of healthy natural enamel. Obtaining a similar hierarchy and cleavage characteristics as natural enamel in the mineralized layer, taking into account the micromorphological features of dental tissue, is an urgent problem for future research. Full article

Atomic deuterium (D) adsorption on free-standing nanoporous graphene obtained by ultra-high vacuum D${}_2$ molecular cracking reveals a homogeneous distribution all over the nanoporous graphene sample, as deduced by ultra-high vacuum Raman spectroscopy combined with core-level photoemission spectroscopy. Raman microscopy unveils the presence of bonding distortion, from the signal associated to the planar sp${}^2$ configuration of graphene toward the sp${}^3$ tetrahedral structure of graphane. The establishment of D–C sp${}^3$ hybrid bonds is also clearly determined by high-resolution X-ray photoelectron spectroscopy and spatially correlated to the Auger spectroscopy signal. This work shows that the low-energy molecular cracking of D${}_2$ in an ultra-high vacuum is an efficient strategy for obtaining high-quality semiconducting graphane with homogeneous uptake of deuterium atoms, as confirmed by this combined optical and electronic spectro-microscopy study wholly carried out in ultra-high vacuum conditions. Full article

The high voltage spinel material LiMn_1.5Ni_0.5O₄ (LMNO) has the potential to increase the energy density of lithium batteries. However, its battery performance suffers from poor long-term cycling and high-temperature stability. In order to overcome these limitations, we have studied the effect of partial substitution of Mn with Ti and LiMn_1.5−x Ni_0.5Ti_xO₄ (x = 0.05, 0.1, 0.3), LMNTO, materials have been synthesized in a newly modified sol-gel method and then characterized by TEM, SEM (EDX), AC Electrochemical Impedance Spectroscopy and Soft X-ray Spectromicroscopy. We have demonstrated that the long-term cycling limitation with these types of materials can be resolved and herein 2000 cycles at a high C-rate have been demonstrated in half cells. We have attributed this behavior to a possible charge compensation mechanism as evidenced by a Soft X-ray Spectromicroscopy study of delithiated LMNTO materials. This work takes high energy density batteries based on high voltage spinel material one step further towards commercialization, and it is believed that further improvement can be achieved using new electrolyte formulations. Full article

In this work, for the first time, the influence of the coordination environment as well as Ca and P atomic states on biomimetic composites integrated with dental tissue was investigated. Bioinspired dental composites were synthesised based on nanocrystalline calcium carbonate-substituted hydroxyapatite $C{a}_4^{\mathrm{I}}C{a}_6^{\mathrm{II}}{\left(P{O}_4\right)}_{6-x}{\left(C{O}_3\right)}_{x+y}{\left(OH\right)}_{2-y}$ (nano-cHAp) obtained from a biogenic source and a set of polar amino acids that modelled the organic matrix. Biomimetic composites, as well as natural dental tissue samples, were investigated using Raman spectromicroscopy and synchrotron X-ray absorption near edge structure (XANES) spectroscopy. Molecular structure and energy structure studies revealed several important features related to the different calcium atomic environments. It was shown that biomimetic composites created in order to reproduce the physicochemical properties of dental tissue provide good imitation of molecular and electron energetic properties, including the carbonate anion CO₃²⁻ and the atomic Ca/P ratio in nanocrystals. The features of the molecular structure of biomimetic composites are inherited from the nano-cHAp (to a greater extent) and the amino acid cocktail used for their creation, and are caused by the ratio between the mineral and organic components, which is similar to the composition of natural enamel and dentine. In this case, violation of the nano-cHAp stoichiometry, which is the mineral basis of the natural and bioinspired composites, as well as the inclusion of different molecular groups in the nano-cHAp lattice, do not affect the coordination environment of phosphorus atoms. The differences observed in the molecular and electron energetic structures of the natural enamel and dentine and the imitation of their properties by biomimetic materials are caused by rearrangement in the local environment of the calcium atoms in the HAp crystal lattice. The surface of the nano-cHAp crystals in the natural enamel and dentine involved in the formation of bonds with the organic matrix is characterised by the coordination environment of the calcium atom, corresponding to its location in the Ca^I position—that is, bound through common oxygen atoms with PO₄ tetrahedrons. At the same time, on the surface of nano-cHAp crystals in bioinspired dental materials, the calcium atom is characteristically located in the Ca^II position, bound to the hydroxyl OH group. The features detected in the atomic and molecular coordination environment in nano-cHAp play a fundamental role in recreating a biomimetic dental composite of the natural organomineral interaction in mineralised tissue and will help to find an optimal way to integrate the dental biocomposite with natural tissue. Full article

Scanning transmission X-ray microscopy (STXM) was utilized for analysing the bioremediation of Cr(VI) by Citrobacter freundii, a species of gram-negative bacteria. The biosorption and bioreduction processes were analysed by the chemical mapping of cells biosorbed at different concentrations of Cr(VI). STXM spectromicroscopy images were recorded at O K-edge and Cr L-edge. A thorough analysis of the X-ray absorption features corresponding to different oxidation states of Cr in the biosorbed cell indicated the coexistence of Cr(III) and Cr(VI) at higher concentrations. This signifies the presence of partially reduced Cr(VI) in addition to biosorbed Cr(VI). In addition, the Cr(III) signal is intense compared with Cr(VI) at different regions of the cell indicating excess of reduced Cr. Speciation of adsorbed Cr was analysed for the spectral features of biosorbed cell and comparison with Cr standards. Analysis of absorption onset, L3/L2 ratio and absorption fine structure concludes that adsorbed Cr is predominantly present as Cr(III) hydroxide or oxyhydroxide. The evolution of absorption features in the duration of biosorption process was also studied. These time lapse studies depict the gradual decrement in Cr(VI) signal as biosorption proceeds. A strong evidence of interaction of Cr with the cell material was also observed. The obtained results provide insights into the biosorption process and chemical speciation of Cr on the cells. Full article