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Animal studies show that exposure to the environmental pollutant 3,3’,4,4’,5-pentachlorobiphenyl (PCB126) causes alterations in hepatic metals as measured in acid-digested volume adjusted tissue. These studies lack the detail of the spatial distribution within the liver. Here we use X-ray fluorescence microscopy (XFM) to assess the spatial distribution of trace elements within liver tissue. Liver samples from male Sprague-Dawley rats, treated either with vehicle or PCB126, were formalin fixed and paraffin embedded. Serial sections were prepared for traditional H&E staining or placed on silicon nitride windows for XFM. With XFM, metal gradients between the portal triad and the central vein were seen, especially with copper and iron. These gradients change with exposure to PCB126, even reverse. This is the first report of how micronutrients vary spatially within the liver, and how they change in response to toxicant exposure. In addition, high concentrations of zinc clusters were discovered in the extracellular space. PCB126-treatment did not affect their presence, but did alter their elemental makeup suggesting a more general biological function. Further work is needed to properly evaluate the gradients and their alterations as well as classify the zinc clusters to determine their role in liver function and zinc homeostasis.

Negative differential resistance behavior in oxide memristors, especially those using NbO2, is gaining renewed interest because of its potential utility in neuromorphic computing. However, there has been a decade-long controversy over whether the negative differential resistance is caused by a relatively low-temperature non-linear transport mechanism or a high-temperature Mott transition. Resolving this issue will enable consistent and robust predictive modeling of this phenomenon for different applications. Here we examine NbO2 memristors that exhibit both a current-controlled and a temperature-controlled negative differential resistance. Through thermal and chemical spectromicroscopy and numerical simulations, we confirm that the former is caused by a ~400 K non-linear-transport-driven instability and the latter is caused by the ~1000 K Mott metal-insulator transition, for which the thermal conductance counter-intuitively decreases in the metallic state relative to the insulating state. The development of future computation devices will be aided by a better understanding of the physics underlying material behaviors. Using thermoreflectance and spatially resolved X-ray microscopy, Kumar et al. elucidate the origin of two types of negative differential resistance in NbO2 memristors.

The past decade has witnessed tremendous growth in both interest and available techniques for laboratory X-ray analysis. From the progression of commercially-available micro- and nano-CT scanners to the resolution and sensitivity enhancements of x-ray fluorescence spectrometers, the scientific community is benefiting from a rapid expansion of laboratory-based x-ray techniques. In our work, we have developed a suite of advanced x-ray instrumentation providing a wide range of enhanced capabilities for specimen characterization. The key enabling technology lies in the X-ray source, which features a microstructured target capable of providing 5-10x higher brightness than conventional sealed-tube x-ray sources and offering power flux densities that rival rotating anode sources. The target array can be custom-designed to incorporate a variety of materials, facilitating fast & easy switching between characteristic emission lines and radiation spectra. This source has been subsequently integrated with state-of-the-art X-ray focusing optics, such as ellipsoidal/paraboloidal capillary lenses and finely-structured Fresnel zone plate imaging objective lenses, and sensitive scintillator-coupled CCD detection systems, opening up new opportunities for advancing laboratory x-ray inspection equipment. Here, we will describe the system geometries in detail and demonstrate how these new advancements have led us to the development of laboratory micro-XRF, nano-XRM, and XAS instrumentation. We will also briefly introduce the image-centric software workspace, which facilitates novice users to collect data quickly and reliably with minimal training overhead.

Talbot-Lau interferometry (TLI) enables X-ray imaging in multiple contrasts, especially for low-Z materials, complementary to conventional absorption radiography. A novel microstructured array anode target (MAAT) source offers major advantages over the combination of an extended X-ray source coupled with an absorption grating (also known as the source grating), including more efficient use of source X-rays and a larger field of view, especially so in the case of Xray energies greater than 30 keV. Therefore, there is an increasing interest in optimizing performance of the MAAT X-ray source to achieve high energy TLI. With the Monte Carlo simulation, here we systematically evaluate the MAAT source parameters for TLI and report the optimal values for high X-ray energies. In particular, the spatial dependence of the Xray generating capability of the MAAT is studied with respect to the incidence angle of the electron beam to the normal of the MAAT target surface.

Biological X-ray fluorescence microscopy (XFM) is an important tool for determining quantitative distributions of bioinorganics and essential trace elements. Here we present a new analysis approach for rapid nanoscale ptychographic imaging and simultaneous chemical mapping of large radiation sensitive specimens without image degradation associated with probe evolution.

CA extern Antidot lattices are potential candidates to act as bit patterned media for data storage as they are able to trap nanoscale magnetic domains between two adjacent holes. Here, we demonstrate the combination of micromagnetic modeling and x-ray microscopy. Detailed simulation of these systems can only be achieved by micromagnetic modeling that takes thermal effects into account. For this purpose, a Landau-Lifshitz-Bloch approach is used here. The calculated melting of magnetic domains within the antidot lattice is reproduced experimentally by x-ray microscopy. Furthermore, we compare conventional scanning transmission x-ray microscopy with resolution enhanced ptychography. Hence, we achieve a resolution of 13 nm. The results demonstrate that ptychographic imaging can also recover magnetic contrast in the presence of a strong topological variation and is generally applicable toward magnetic samples requiring ultimate resolution. © 2020 Author(s).

Summary Trace metals play important roles in biological function, and x-ray fluorescence microscopy (XFM) provides a way to quantitatively image their distribution within cells. The faithfulness of these measurements is dependent on proper sample preparation. Using mouse embryonic fibroblast NIH/3T3 cells as an example, we compare various approaches to the preparation of adherent mammalian cells for XFM imaging under ambient temperature. Direct side-by-side comparison shows that plunge-freezing-based cryoimmobilization provides more faithful preservation than conventional chemical fixation for most biologically important elements including P, S, Cl, K, Fe, Cu, Zn and possibly Ca in adherent mammalian cells. Although cells rinsed with fresh media had a great deal of extracellular background signal for Cl and Ca, this approach maintained cells at the best possible physiological status before rapid freezing and it does not interfere with XFM analysis of other elements. If chemical fixation has to be chosen, the combination of 3% paraformaldehyde and 1.5 % glutaraldehyde preserves S, Fe, Cu and Zn better than either fixative alone. When chemically fixed cells were subjected to a variety of dehydration processes, air drying was proved to be more suitable than other drying methods such as graded ethanol dehydration and freeze drying. This first detailed comparison for x-ray fluorescence microscopy shows how detailed quantitative conclusions can be affected by the choice of cell preparation method.

Owing to its extreme sensitivity, quantitative mapping of elemental distributionsviaX-ray fluorescence microscopy (XFM) has become a key microanalytical technique. The recent realisation of scanning X-ray diffraction microscopy (SXDM) meanwhile provides an avenue for quantitative super-resolved ultra-structural visualization. The similarity of their experimental geometries indicates excellent prospects for simultaneous acquisition. Here, in both step- and fly-scanning modes, robust, simultaneous XFM-SXDM is demonstrated.

Talbot-Lau interferometry (TLI) provides additional contrast modes for x-ray imaging that are complementary to conventional absorption radiography. TLI is particularly interesting because it is one of the few practical methods for realizing phase contrast with x-rays that is compatible with large-spot high power x-ray sources. A novel micro array anode structured target (MAAST) x-ray source offers several advantages for TLI over the conventional combination of an extended x-ray source coupled with an absorption grating including higher flux and larger field of view, and these advantages become more pronounced for x-ray energies in excess of 30 keV. A Monte Carlo simulation was performed to determine the optimal parameters for a MAAST source for use with TLI. It was found that the both spatial distribution of x-ray production and the number of x-ray produced in the MAAST have a strong dependence on the incidence angle of the electron beam.

Lithium-rich layered transition metal oxide positive electrodes offer access to anion redox at high potentials, thereby promising high energy densities for lithium-ion batteries. However, anion redox is also associated with several unfavorable electrochemical properties, such as open-circuit voltage hysteresis. Here we reveal that in Li1.17–xNi0.21Co0.08Mn0.54O2, these properties arise from a strong coupling between anion redox and cation migration. We combine various X-ray spectroscopic, microscopic, and structural probes to show that partially reversible transition metal migration decreases the potential of the bulk oxygen redox couple by > 1 V, leading to a reordering in the anionic and cationic redox potentials during cycling. First principles calculations show that this is due to the drastic change in the local oxygen coordination environments associated with the transition metal migration. We propose that this mechanism is involved in stabilizing the oxygen redox couple, which we observe spectroscopically to persist for 500 charge/discharge cycles., Lithium ion battery electrodes employing anion redox exhibit high energy densities but suffer from poor cyclability. Here the authors reveal that the voltage of anion redox is strongly affected by structural changes that occur during battery cycling, explaining its unique electrochemical properties.

Further improvements in the low frequency sensitivity of gravitational wave detectors are important for increasing the observable population of astrophysical sources, such as intermediate mass compact black hole binary systems. Improvements in the lower stage mirror and suspension systems will set challenging targets for the required thermal noise performance of the cantilever blade springs, which provide vertical softness and, thus, isolation to the mirror suspension stack. This is required due to the coupling between the vertical and horizontal axes due to the curvature of the Earth. This can be achieved through use of high mechanical Q materials, which are compatible with cryogenic cooling, such as crystalline silicon. However, such materials are brittle, posing further challenges for assembly/jointing and, more generally, for long-term robustness. Here, we report on experimental studies of the breaking strength of silicon at room temperature, via both tensile and 4-point flexural testing; and on the effects of various surface treatments and coatings on durability and strength. Single- and multi-layer DLC (diamond-like carbon) coatings, together with magnetron-sputtered silica and thermally-grown silica, are investigated, as are the effects of substrate preparation and argon plasma pre-treatment. Application of single- or multi-layer DLC coatings can significantly improve the failure stress of silicon flexures, in addition to improved robustness for handling (assessed through abrasion tests). Improvements of up to 80% in tensile strength, a twofold increase in flexural strength, in addition to a 6.4 times reduction in the vertical thermal noise contribution of the suspension stack at 10 Hz are reported (compared to current Advanced LIGO design). The use of silicon blade springs would also significantly reduce potential 'crackling noise' associated with the underlying discrete events associated with plastic deformation in loaded flexures.

Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolution beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and ∼90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. This combined approach offers a way to study the role of trace elements in their structural context.

© the Owner Societies 2015.Lithium ion batteries are the dominant form of energy storage in mobile devices, increasingly employed in transportation, and likely candidates for renewable energy storage and integration into the electrical grid. To fulfil their powerful potential, electrodes with increased capacity, faster charge rates, and longer cycle life must be developed. Understanding the mechanics and chemistry of individual nanoparticles under in situ conditions is a crucial step to improving performance and mitigating damage. Here we reveal three-dimensional strain evolution within a single nanoparticle of a promising high voltage cathode material, LiNi0.5Mn1.5O4, under in situ conditions. The particle becomes disconnected during the second charging cycle. This is attributed to the formation of a cathode electrolyte interphase layer with slow ionic conduction. The three-dimensional strain pattern within the particle is independent of cell voltage after disconnection, indicating that the particle is unable to redistribute lithium within its volume or to its neighbours. Understanding the disconnection process at the single particle level and the equilibrium or non-equilibrium state of nanoparticles is essential to improving performance of current and future electrochemical energy storage systems.

Motivated by the Advanced Photon Source Upgrade (APS-U), a new hard X-ray microscope called “Velociprobe” has been recently designed and built for fast ptychographic imaging with high spatial resolution. We are addressing the challenges of high-resolution and fast scanning with novel hardware/stage designs, new positioner control designs, and new data acquisition strategies, including the use of high bandwidth interferometric measurements. The use of granite, air-bearing-supported stages provides the necessary long travel ranges for coarse motion to accommodate real samples and variable energy operation while remaining highly stable during fine scanning. Scanning the low-mass zone plate enables high-speed high-precision motion of the probe over the sample. Our primary goal is to use this instrument to demonstrate sub-10 nm spatial resolution ptychography over a 1-square-micron area in under 10 seconds. We have also designed the instrument to take advantage of the upgraded source when the APS-U is completed. This presentation will describe the unique designs and characteristics of this instrument, and some preliminary data obtained during the instrument commission.

X-ray microscopy can be used to image whole, unsectioned cells in their native hydrated state. It complements the higher resolution of electron microscopy for submicrometer thick specimens, and the molecule-specific imaging capabilites of fluorescence light microscopy. We describe here the first use of fast, continuous x-ray scanning of frozen hydrated cells for simultaneous sub-20 nm resolution ptychographic transmission imaging with high contrast, and sub-100 nm resolution deconvolved x-ray fluorescence imaging of diffusible and bound ions at native concentrations, without the need to add specific labels. By working with cells that have been rapidly frozen without the use of chemical fixatives, and imaging them under cryogenic conditions, we are able to obtain images with well preserved structural and chemical composition, and sufficient stability against radiation damage to allow for multiple images to be obtained with no observable change.

While the recent establishment of the role of thermophoresis/diffusion-driven oxygen migration during resistance switching in metal oxide memristors provided critical insights required for memristor modeling, extended investigations of the role of oxygen migration during ageing and failure remain to be detailed. Such detailing will enable failure-tolerant design, which can lead to enhanced performance of memristor-based next-generation storage-class memory. Here we directly observed lateral oxygen migration using in-situ synchrotron x-ray absorption spectromicroscopy of HfOx memristors during initial resistance switching, wear over millions of switching cycles, and eventual failure, through which we determined potential physical causes of failure. Using this information, we reengineered devices to mitigate three failure mechanisms, and demonstrated an improvement in endurance of about three orders of magnitude., Comment: 5 pages, 4 figures

The role of ions in the fungal decay process of lignocellulose biomaterials, and more broadly fungal metabolism, has implications for diverse research disciplines ranging from plant pathology and forest ecology, to carbon sequestration. Despite the importance of ions in fungal decay mechanisms, the spatial distribution and quantification of ions in lignocellulosic cell walls and fungal hyphae during decay is not known. Here we employ synchrotron-based X-ray fluorescence microscopy (XFM) to map and quantify physiologically relevant ions, such as K, Ca, Mn, Fe, and Zn, in wood being decayed by the model brown rot fungus Serpula lacrymans. Two-dimensional XFM maps were obtained to study the ion spatial distributions from mm to submicron length scales in wood, fungal hyphae with the dried extracellular matrix (ECM) from the fungus, and Ca oxalate crystals. Three-dimensional ion volume reconstructions were also acquired of wood cell walls and hyphae with ECM. Results show that the fungus actively transports some ions, such as Fe, into the wood and controls the distribution of ions at both the bulk wood and cell wall length scales. These measurements provide new insights into the movement of ions during decay and illustrate how synchrotron-based XFM is uniquely suited study these ions.

Data Exchange is a simple data model designed to interface, or `exchange', data among different instruments, and to enable sharing of data analysis tools. Data Exchange focuses on technique rather than instrument descriptions, and on provenance tracking of analysis steps and results. In this paper the successful application of the Data Exchange model to a variety of X-ray techniques, including tomography, fluorescence spectroscopy, fluorescence tomography and photon correlation spectroscopy, is described.

Phase-diverse X-ray coherent diffractive imaging (CDI) provides a route to high sensitivity and spatial resolution with moderate radiation dose. It also provides a robust solution to the well-known phase-problem, making on-line image reconstruction feasible. Here we apply phase-diverse CDI to a cellular sample, obtaining images of an erythrocyte infected by the sexual stage of the malaria parasite, Plasmodium falciparum, with a radiation dose significantly lower than the lowest dose previously reported for cellular imaging using CDI. The high sensitivity and resolution allow key biological features to be identified within intact cells, providing complementary information to optical and electron microscopy. This high throughput method could be used for fast tomographic imaging, or to generate multiple replicates in two-dimensions of hydrated biological systems without freezing or fixing. This work demonstrates that phase-diverse CDI is a valuable complementary imaging method for the biological sciences and ready for immediate application.

Biological productivity in many ocean regions is controlled by the availability of the nutrient phosphorus. In the Mediterranean Sea, aerosol deposition is a key source of phosphorus and understanding its composition is critical for determining its potential bioavailability. Aerosol phosphorus was investigated in European and North African air masses using phosphorus near-edge X-ray fluorescence spectroscopy (P-NEXFS). These air masses are the main source of aerosol deposition to the Mediterranean Sea. We show that European aerosols are a significant source of soluble phosphorus to the Mediterranean Sea. European aerosols deliver on average 3.5 times more soluble phosphorus than North African aerosols and furthermore are dominated by organic phosphorus compounds. The ultimate source of organic phosphorus does not stem from common primary emission sources. Rather, phosphorus associated with bacteria best explains the presence of organic phosphorus in Mediterranean aerosols.

Transition metal oxide memristors, or resistive random-access memory (RRAM) switches, are under intense development for storage-class memory because of their favorable operating power, endurance, speed, and density. Their commercial deployment critically depends on predictive compact models based on understanding nanoscale physico-chemical forces, which remains elusive and controversial owing to the difficulties in directly observing atomic motions during resistive switching, Here, using scanning transmission synchrotron x-ray spectromicroscopy to study in-situ switching of hafnium oxide memristors, we directly observed the formation of a localized oxygen-deficiency-derived conductive channel surrounded by a low-conductivity ring of excess oxygen. Subsequent thermal annealing homogenized the segregated oxygen, resetting the cells towards their as-grown resistance state. We show that the formation and dissolution of the conduction channel are successfully modeled by radial thermophoresis and Fick diffusion of oxygen atoms driven by Joule heating. This confirmation and quantification of two opposing nanoscale radial forces that affect bipolar memristor switching are important components for any future physics-based compact model for the electronic switching of these devices., Comment: 6 pages and 4 figures

We analyzed micrometer-scale titanium-niobium-oxide prototype memristors, which exhibited low write-power (millions of cycles). To understand their physico-chemical operating mechanisms, we performed in-operando synchrotron x-ray transmission nanoscale spectromicroscopy using an ultra-sensitive time-multiplexed technique. We observed only spatially uniform material changes during cell operation, in sharp contrast to the frequently detected formation of a localized conduction channel in transition-metal-oxide memristors. We also associated the response of assigned spectral features distinctly to non-volatile storage (resistance change) and writing of information (application of voltage and Joule heating). These results provide critical insights into high-performance memristors that will aid in device design, scaling and predictive circuit-modeling, all of which are essential for the widespread deployment of successful memristor applications., 5 pages 4 fgures

Motivated by the advanced photon source upgrade, a new hard X-ray microscope called "Velociprobe" has been recently designed and built for fast ptychographic imaging with high spatial resolution. We are addressing the challenges of high-resolution and fast scanning with novel hardware designs, advanced motion controls, and new data acquisition strategies, including the use of high-bandwidth interferometric measurements. The use of granite, air-bearing-supported stages provides the necessary long travel ranges for coarse motion to accommodate real samples and variable energy operation while remaining highly stable during fine scanning. Scanning the low-mass zone plate enables high-speed and high-precision motion of the probe over the sample. With an advanced control algorithm implemented in a closed-loop feedback system, the setup achieves a position resolution (3σ) of 2 nm. The instrument performance is evaluated by 2D fly-scan ptychography with our developed data acquisition strategies. A spatial resolution of 8.8 nm has been demonstrated on a Au test sample with a detector continuous frame rate of 200 Hz. Using a higher flux X-ray source provided by double-multilayer monochromator, we achieve 10 nm resolution for an integrated circuit sample in an ultrafast scan with a detector's full continuous frame rate of 3000 Hz (0.33 ms per exposure), resulting in an outstanding imaging rate of 9 × 104 resolution elements per second.

Talbot-Lau grating interferometer (TLGI) has great advantages in X-ray imaging contrasts, especially for low-Z materials, over conventional absorption contrast. A microstructured array anode target (MAAT) source offers significantly higher imaging throughput than the combination of an extended X-ray source paired with an absorption grating (also known as source grating). The performance of the MAAT source can be optimized with respect to the areal density, dimensions, and choice of material for the microstructured metal inserts (MMI) and the substrate in which they are embedded. In this paper, we analyze the X-ray generation efficiency per incident electron, relative fraction of X-rays generated by MMI and substrate, X-ray spectrum, and angular distribution via Monte Carlo simulation. Based on the simulation results, the optimal parameters are obtained for a MAAT with incident electron energies from 30 keV to 120 keV. The corresponding temperature distribution within the MAAT is also simulated for the optimal set of the parameters via finite element analysis. As demonstrated by the thermal analysis data, the maximum allowable electron-beam power loading was derived that allows a stable operation of the transmission MAAT.

The performance of a diamond X-ray phase retarder to enable the production of circularly polarized X-rays has been quantitatively measured and magnetic dichroism contrast in transmission and diffraction geometries has been demonstrated. Feasibility tests for dichroic Bragg coherent diffractive imaging experiments were performed and showed that the diamond X-ray phase retarder does not produce significant distortions to the X-ray wavefront and that Bragg coherent diffractive imaging reconstructions are achievable., Recent progress in the development of dichroic Bragg coherent diffractive imaging, a new technique for simultaneous three-dimensional imaging of strain and magnetization at the nanoscale, is reported. This progress includes the installation of a diamond X-ray phase retarder at beamline 34-ID-C of the Advanced Photon Source. The performance of the phase retarder for tuning X-ray polarization is demonstrated with temperature-dependent X-ray magnetic circular dichroism measurements on a gadolinium foil in transmission and on a Gd5Si2Ge2 crystal in diffraction geometry with a partially coherent, focused X-ray beam. Feasibility tests for dichroic Bragg coherent diffractive imaging are presented. These tests include (1) using conventional Bragg coherent diffractive imaging to determine whether the phase retarder introduces aberrations using a nonmagnetic gold nanocrystal as a control sample, and (2) collecting coherent diffraction patterns of a magnetic Gd5Si2Ge2 nanocrystal with left- and right-circularly polarized X-rays. Future applications of dichroic Bragg coherent diffractive imaging for the correlation of strain and lattice defects with magnetic ordering and inhomogeneities are considered.

Journal of synchrotron radiation 24(1), 83 - 94(2017). doi:10.1107/S1600577516017525, The proliferation of extremely intense synchrotron sources has enabled ever higher-resolution structures to be obtained using data collected from smaller and often more imperfect biological crystals (Helliwell, 1984). Synchrotron beamlines now exist that are capable of measuring data from single crystals that are just a few micrometres in size. This provides renewed motivation to study and understand the radiation damage behaviour of small protein crystals. Reciprocal-space mapping and Bragg coherent diffractive imaging experiments have been performed on cryo-cooled microcrystals of hen egg-white lysozyme as they undergo radiation damage. Several well established metrics, such as intensity-loss and lattice expansion, are applied to the diffraction data and the results are compared with several new metrics that can be extracted from the coherent imaging experiments. Individually some of these metrics are inconclusive. However, combining metrics, the results suggest that radiation damage behaviour in protein micro-crystals differs from that of larger protein crystals and may allow them to continue to diffract for longer. A possible mechanism to account for these observations is proposed., Published by IUCr, Chester

The chemical and physical speciation of atmospheric sulfur was investigated in ambient aerosol samples using a combination of sulfur near-edge x-ray fluorescence spectroscopy (S-NEXFS) and X-ray fluorescence (XRF) microscopy. These techniques were used to determine the composition and oxidation state of sulfur in common primary emission sources and ambient particulate matter collected from the greater Atlanta area. Ambient particulate matter samples contained two oxidation states: S0 and S+VI. Ninety-five percent of the individual aerosol particles (> 1 µm) analyzed contain S0. Linear combination fitting revealed that S+VI in ambient aerosol was dominated by ammonium sulfate as well as metal sulfates. The finding of metal sulfates provides further evidence for acidic reactions that solubilize metals, such as iron, during atmospheric transport. Emission sources, including biomass burning, coal fly ash, gasoline, diesel, volcanic ash, and aerosolized Atlanta soil, and the commercially available bacterium Bacillus subtilis, contained only S+VI. A commercially available Azotobacter vinelandii sample contained approximately equal proportions of S0 and S+VI. S0 in individual aerosol particles most likely originates from primary emission sources, such as aerosolized bacteria or incomplete combustion.

X-ray fluorescence offers unparalleled sensitivity for imaging the nanoscale distribution of trace elements in micrometer thick samples, while x-ray ptychography offers an approach to image light element containing structures at a resolution beyond that of the x-ray lens used. These methods can be used in combination, and in continuous scan mode for rapid data acquisition when using multiple probe mode reconstruction methods. We discuss here the opportunities and limitations of making use of additional information provided by ptychography to improve x-ray fluorescence images in two ways: by using position-error-correction algorithms to correct for scan distortions in fluorescence scans, and by considering the signal-to-noise limits on previously-demonstrated ptychographic probe deconvolution methods. This highlights the advantages of using a combined approach.

Trace elements, particularly metals, play very important roles in biological systems. Synchrotron-based hard X-ray fluorescence microscopy offers the most suitable capabilities to quantitatively study trace metals in thick biological samples, such as whole cells and tissues. In this manuscript, we have demonstrated X-ray fluorescence imaging of frozen-hydrated whole cells using the recent developed Bionanoprobe (BNP). The BNP provides spatial resolution down to 30 nm and cryogenic capabilities. Frozen-hydrated biological cells have been directly examined on a sub-cellular level at liquid nitrogen temperatures with minimal sample preparation.