Gareth Moorhead

The X‐ray Fluorescence Microscopy Beamline at the Australian Synchrotron

A hard x‐ray micro‐nanoprobe has commenced operation at the Australian Synchrotron providing versatile x‐ray fluorescence microscopy across an incident energy range from 4 to 25 keV. Two x‐ray probes are used to collect μ‐XRF and μ‐XANES for elemental and chemical microanalysis: a Kirkpatrick‐Baez mirror microprobe for micron resolution studies and a Fresnel zone plate nanoprobe capable of 60‐nm resolution. Some unique aspects of the beamline design and operation are discussed. An advanced energy dispersive x‐ray fluorescence detection scheme named Maia has been developed for the beamline, which enables ultrafast x‐ray fluorescence microscopy.

The Maia Spectroscopy Detector System: Engineering for Integrated Pulse Capture, Low-Latency Scanning and Real-Time Processing

The Maia detector system is engineered for energy dispersive x‐ray fluorescence spectroscopy and elemental imaging at photon rates exceeding 107/s, integrated scanning of samples for pixel transit times as small as 50μs and high definition images of 108 pixels and real‐time processing of detected events for spectral deconvolution and online display of pure elemental images. The system developed by CSIRO and BNL combines a planar silicon 384 detector array, application‐specific integrated circuits for pulse shaping and peak detection and sampling and optical data transmission to an FPGA‐based pipelined, parallel processor. This paper describes the system and the underpinning engineering solutions.

Quantification of ZnO Nanoparticle Uptake, Distribution, and Dissolution within Individual Human Macrophages

The usefulness of zinc oxide (ZnO) nanoparticles has led to their wide distribution in consumer products, despite only a limited understanding of how this nanomaterial behaves within biological systems. From a nanotoxicological viewpoint the interaction(s) of ZnO nanoparticles with cells of the immune system is of specific interest, as these nanostructures are readily phagocytosed. In this study, rapid scanning X-ray fluorescence microscopy was used to assay the number ZnO nanoparticles associated with ∼1000 individual THP-1 monocyte-derived human macrophages. These data showed that nanoparticle-treated cells endured a 400% elevation in total Zn levels, 13-fold greater than the increase observed when incubated in the presence of an equitoxic concentration of ZnCl2. Even after excluding the contribution of internalized nanoparticles, Zn levels in nanoparticle treated cells were raised ∼200% above basal levels. As dissolution of ZnO nanoparticles is critical to their cytotoxic response, we utilized a strategy combining ion beam milling, X-ray fluorescence and scanning electron microscopy to directly probe the distribution and composition of ZnO nanoparticles throughout the cellular interior. This study demonstrated that correlative photon and ion beam imaging techniques can provide both high-resolution and statistically powerful information on the biology of metal oxide nanoparticles at the single-cell level. Our approach promises ready application to broader studies of phenomena at the interface of nanotechnology and biology.

Maia X-ray fluorescence imaging: Capturing detail in complex natural samples

Motivated by the challenge of capturing complex hierarchical chemical detail in natural material from a wide range of applications, the Maia detector array and integrated realtime processor have been developed to acquire X-ray fluorescence images using X-ray Fluorescence Microscopy (XFM). Maia has been deployed initially at the XFM beamline at the Australian Synchrotron and more recently, demonstrating improvements in energy resolution, at the P06 beamline at Petra III in Germany. Maia captures fine detail in element images beyond 100 M pixels. It combines a large solid-angle annular energy-dispersive 384 detector array, stage encoder and flux counter inputs and dedicated FPGA-based real-time event processor with embedded spectral deconvolution. This enables high definition imaging and enhanced trace element sensitivity to capture complex trace element textures and place them in a detailed spatial context. Maia hardware and software methods provide per pixel correction for dwell, beam flux variation, dead-time and pileup, as well as off-line parallel processing for enhanced throughput. Methods have been developed for real-time display of deconvoluted SXRF element images, depth mapping of rare particles and the acquisition of 3D datasets for fluorescence tomography and XANES imaging using a spectral deconvolution method that tracks beam energy variation.

High-Definition X-ray Fluorescence Elemental Mapping of Paintings

A historical self-portrait painted by Sir Arthur Streeton (1867-1943) has been studied with fast-scanning X-ray fluorescence microscopy using synchrotron radiation. One of the techniques unique strengths is the ability to reveal metal distributions in the pigments of underlying brushstrokes, thus providing information critical to the interpretation of a painting. We have applied the nondestructive technique with the event-mode Maia X-ray detector, which has the capability to record elemental maps at megapixels per hour with the full X-ray fluorescence spectrum collected per pixel. The painting poses a difficult challenge to conventional X-ray analysis, because it was completely obscured with heavy brushstrokes of highly X-ray absorptive lead white paint (2PbCO(3)·Pb(OH)(2)) by the artist, making it an excellent candidate for the application of the synchrotron-based technique. The 25 megapixel elemental maps were successfully observed through the lead white paint across the 200 × 300 mm(2) scan area. The sweeping brushstrokes of the lead white overpaint contributed significant detrimental structure to the elemental maps. A corrective procedure was devised to enhance the visualization of the elemental maps by using the elastic X-ray scatter as a proxy for the lead white overpaint. We foresee the technique applied to the most demanding of culturally significant artworks where conventional analytical methods are inadequate.

Quantitative comparison of preparation methodologies for x-ray fluorescence microscopy of brain tissue

X-ray fluorescence microscopy (XFM) facilitates high-sensitivity quantitative imaging of trace metals at high spatial resolution over large sample areas and can be applied to a diverse range of biological samples. Accurate determination of elemental content from recorded spectra requires proper calibration of the XFM instrument under the relevant operating conditions. Here, we describe the manufacture, characterization, and utilization of multi-element thin-film reference foils for use in calibration of XFM measurements of biological and other specimens. We have used these internal standards to assess the two-dimensional distribution of trace metals in a thin tissue section of a rat hippocampus. The data used in this study was acquired at the XFM beamline of the Australian Synchrotron using a new 384-element array detector (Maia) and at beamline 2-ID-E at the Advanced Photon Source. Post-processing of samples by different fixation techniques was investigated, with the conclusion that differences in solvent type and sample handling can significantly alter elemental content. The present study highlights the quantitative capability, high statistical power, and versatility of the XFM technique for mapping trace metals in biological samples, e.g., brain tissue samples in order to help understand neurological processes, especially when implemented in conjunction with a high-performance detector such as Maia.

Caenorhabditis elegans Maintains Highly Compartmentalized Cellular Distribution of Metals and Steep Concentration Gradients of Manganese

Bioinorganic chemistry is critical to cellular function. Homeostasis of manganese (Mn), for example, is essential for life. A lack of methods for direct in situ visualization of Mn and other biological metals within intact multicellular eukaryotes limits our understanding of management of these metals. We provide the first quantitative subcellular visualization of endogenous Mn concentrations (spanning two orders of magnitude) associated with individual cells of the nematode, Caenorhabditis elegans.

Maia X-ray Microprobe Detector Array System

Maia is an advanced system designed specifically for scanning x-ray fluorescence microprobe applications. It consists of a large array of photodiode detectors and associated signal processing, closely coupled to an FPGA-based control and analysis system. In this paper we will describe the architecture and construction of the system.

Evaluation of p-stop structures in the n-side of n-on-n silicon strip detectors

A large area (63.6 mm/spl times/64 mm) n-on-n silicon strip detector was fabricated, implementing various p-stop structures in the n-side. The detectors were characterized in laboratory and in beam tests. The inter-strip capacitance showed features in which the individual p-stop structure had the longest tail toward saturation. The beam tests showed other p-stop structures collected more charge in the mid-strip region than the individual p-stop structure. In addition, there was a source which lost or spread charge and induced noise where the over-depletion was insufficient.

The Maia detector array and x-ray fluorescence imaging system: locating rare precious metal phases in complex samples

X-ray fluorescence images acquired using the Maia large solid-angle detector array and integrated real-time processor on the X-ray Fluorescence Microscopy (XFM) beamline at the Australian Synchrotron capture fine detail in complex natural samples with images beyond 100M pixels. Quantitative methods permit real-time display of deconvoluted element images and for the acquisition of large area XFM images and 3D datasets for fluorescence tomography and chemical state (XANES) imaging. This paper outlines the Maia system and analytical methods and describes the use of the large detector array, with a wide range of X-ray take-off angles, to provide sensitivity to the depth of features, which is used to provide an imaging depth contrast and to determine the depth of rare precious metal particles in complex geological samples.