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Near-ambient pressure XPS makes it possible to characterise samples not compatible to ultra-high vacuum, and enables the study of liquid-solid, gas-liquid and gas-solid interfaces. NAP-XPS meas-urements of biofilms, suspended nanoparticles and metal-organic frameworks were performed with EnviroESCA developed by SPECS.
An interesting application is surface characterisation of biofilms, which are bacterial communities embedded in a self-produced polysaccharide matrix. Various model systems ranging from pure polysaccharides of alginate to biofilms harvested directly from the growth medium have been char-acterised in humid conditions[1].
NAP-XPS also makes it possible to characterise nanoparticles in solution. Silver nanoparticles in aqueous solution were characterised and the Ag 3d-spectrum compared to spectra obtained of dried nanoparticles in UHV-conditions[2]. The binding energy of the Ag 3d-core level peak was shifted by 0,6 eV towards higher binding energy for suspended nanoparticles compared to the dried sample measured in UHV. This can be assigned to a change in surface potential at the water-nanoparticle interface.
Metal-organic frameworks (MOFs) are suitable materials for gas storage of small molecules due to their nanoporous, crystalline structure. However, instability in humidity remains an issue for many types of MOFs. XPS-measurements of the MOF-structure HKUST-1 were performed in various NAP-conditions to assess the stability of the sample and its interaction with the gas molecules as water, methanol and pyridine.
Synchrotron XPS in the soft-X-ray regime is suitable for the detection of light elements commonly found in biological samples. Various model systems of biofilms have been developed and characterised at synchrotron- and lab-based facilities. By obtaining the chemical composition at various information depths, the vertical distribution of iodine in an artificial biofilm have been determined.
Bacterial samples are typically freeze dried or cryo-prepared prior to XPS analysis to allow for measurements in ultra-high vacuum (UHV). The sample environment in the near-ambient pressure (NAP) XPS instrument EnviroESCA allows for measurements in up to 15 mbar water vapor, thus, sample preparation is no longer restricted to UHV-compatible techniques. For instance, biofilms grown in medium can be transferred directly from the medium to the measurements chamber, maintaining a humid environment throughout the measurements. Considering the complexity of bacterial samples, sample preparation must be carefully considered in order to obtain meaningful and reproducible results.
In this talk, various strategies for sample preparation of bacteria and biofilms for NAP-XPS measurements will be discussed. Model systems of planktonic bacteria, artificial biofilms resembling the exopolysaccharide matrix and biofilms have been characterised in various conditions. The stability and homogeneity of the samples was assessed by monitoring the C1s core level peak at different sample locations. The quality of the XPS-spectra is also influenced by the gas environment, which will be exemplified by core level spectra of P. Fluorescens acquired in air, water vapor and ultra-high vacuum.
Microbiologically Influenced Corrosion of Cast Iron Containers for High-Level Nuclear Waste Disposal
(2022)
Ductile and corrosion resistant cast iron is investigated as a potential container material to store high-level nuclear waste (HLW) in deep geological repositories (DGR) in claystone bedrock. The dynamic corrosion process is dependent on the conditions present in the DGR which are influenced and/or controlled by geochemical parameters (e.g., redox potential, pH, presence of and ionic concentration in (pore-)water), physical parameters (e.g., pressure), and the influence of metabolically active microorganisms. Cast iron corrosion will occur at the intersection of container and its decontaminable coating with the bentonite backfill material which contains natural microbial populations.
The conditions in a DGR are simulated in microcosm experiments to investigate the impact of microbiologically influenced corrosion (MIC); the microcosms contain: B27 bentonite, synthetic pore water, N2 or N2-CO2 atmosphere, cast iron coupons, as well as the bacterium Desulfosporosinus burensis (isolated from repository depth in Buré, France). Three coupon configurations will be used: untreated, coated with decontaminable coating, and coated with decontaminable coating which has been damaged to simulate possible damages. The microcosms will be examined for bio- and geochemical parameters, such as pH, redox potential, mineral phases, sulphate concentration, Fe(II):Fe(III), changes in microbial populations, and the corrosion process for formation of corrosion products, and potential microbial influence, after a 270-day incubation period at 25°C under anaerobic conditions. In subsequent experiments, the sorption behavior of lanthanides and actinides onto the membranes of viable cells and spores of D. burensis, as well as the surface of corroded cast iron coupons will be investigated.
Maintaining and modulating mechanical anisotropy is essential for biological processes. However, how this is achieved at the microscopic scale in living soft matter is not always clear. Although Brillouin light scattering (BLS) spectroscopy can probe the mechanical properties of materials, spatiotemporal mapping of mechanical anisotropies in living matter with BLS microscopy has been complicated by the need for sequential measurements with tilted excitation and detection angles. Here we introduce Brillouin light scattering anisotropy microscopy (BLAM) for mapping high-frequency viscoelastic anisotropy inside living cells. BLAM employs a radial virtually imaged phased array that enables the collection of angle-resolved dispersion in a single shot, thus enabling us to probe phonon modes in living matter along different directions simultaneously. We demonstrate a precision of 10 MHz in the determination of the Brillouin frequency shift, at a spatial resolution of 2 µm. Following proof-of-principle experiments on muscle myofibres, we apply BLAM to the study of two fundamental biological processes. In plant cell walls, we observe a switch from anisotropic to isotropic wall properties that may lead to asymmetric growth. In mammalian cell nuclei, we uncover a spatiotemporally oscillating elastic anisotropy correlated to chromatin condensation. Our results highlight the role that high-frequency mechanics can play in the regulation of diverse fundamental processes in biological systems. We expect BLAM to find diverse applications in biomedical imaging and material characterization.
This study details a theoretical analysis of leaky and waveguide modes in biperiodic all-dielectric holograms. By tuning diffraction orders and subsequently confining local density of optical states at two distinct resonance wavelengths, we present a new class of highly sensitive refractive index biosensing platforms that are capable of resolving 35.5 to 41.3 nm/RIU of spectral shift for two separate biological analytes.
This study details a thorough analysis of leaky and waveguide modes in biperiodic diffractive nanostructures. By tuning diffraction orders and subsequently confining local density of optical states at two distinct resonance wavelengths, we present a highly sensitive refractive index biosensing platform that can resolve 35.5 to 41.3 nm/RIU of spectral shift for two separate biological analytes.
Consequences of tolerance to disinfectants on the evolution of antibiotic resistance in E. coli
(2023)
Biocides are used as disinfectants and preservatives; one important active substance in biocides is benzalkonium chloride (BAC). BAC-tolerant bacterial strains can survive short treatments with high concentrations of BAC. BAC tolerance and resistance have been linked to antibiotic resistance. Here, the selection dynamics between a BAC-tolerant Escherichia coli strain and a sensitive wild type were investigated under four conditions: in the absence of antibiotics and in the presence of three different sub-inhibitory concentrations of the antibiotic ciprofloxacin in liquid cultures. The wild type was selected over the BAC-tolerant strain in the absence of antibiotics, while the BAC-tolerant strain was selected over the wild type at all ciprofloxacin concentrations investigated, with a minimum selection concentration (MSC) of 1/10th of the minimum inhibitory concentration (MIC) of the wild type. Furthermore, the evolvability of resistance of the two strains to inhibitory concentrations of ciprofloxacin was assessed by performing a serial dilution evolution experiment with gradually increasing ciprofloxacin concentrations. The wild type had a higher probability to develop resistance
to ciprofloxacin than the tolerant strain. By the end of the evolution experiment both strains evolved to grow at the highest ciprofloxacin concentration investigated, which was 2048 ×MIC of the wild type. The importance of these results is highlighted by the fact that concentrations of ciprofloxacin well above the calculated MSC can be found in environmental samples such as hospital wastewaters and livestock slurry. In turn, BAC is used as a disinfectant in the same settings. Thus, the selection of BAC-tolerant strains at sub-inhibitory concentrations of ciprofloxacin can contribute to the stabilization and spread of BAC-tolerance in natural populations. The prevalence of such strains can impair the effects of BAC disinfections.
We investigated the selection dynamics between a benzalkonium chloride (BAC)-tolerant Escherichia coli strain (S4) and a sensitive wild type under four conditions: in the absence of antibiotics and in the presence of three different sub-inhibitory concentrations of the antibiotic ciprofloxacin in liquid cultures. The wild type was selected over the BAC-tolerant strain in the absence of antibiotics, while the opposite was observed at all ciprofloxacin concentrations investigated.Furthermore, we assessed the evolvability of resistance of the two strains to inhibitory concentrations of ciprofloxacin by performing a serial dilution evolution experiment with gradually increasing ciprofloxacin concentrations. The wild type had a higher probability to develop resistance to ciprofloxacin than the tolerant strain. By the end of the evolution experiment both strains evolved to grow at the highest ciprofloxacin concentration investigated, which was 2048 ×MIC of the wild type.