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Over the last decade ptychography has progressed rapidly from a specialist ultramicroscopy technique into a mature method accessible to non‐expert users. However, to improve scientific value ptychography data must reconstruct reliably, with high image quality and at no cost to other correlative methods. Presented here is the implementation of high‐speed ptychography used at the Australian Synchrotron on the XFM beamline, which includes a free‐run data collection mode where dead time is eliminated and the scan time is optimized. It is shown that free‐run data collection is viable for fast and high‐quality ptychography by demonstrating extremely high data rate acquisition covering areas up to 352 000 µm2 at up to 140 µm2 s−1, with 13× spatial resolution enhancement compared with the beam size. With these improvements, ptychography at velocities up to 250 µm s−1 is approaching speeds compatible with fast‐scanning X‐ray fluorescence microscopy. The combination of these methods provides morphological context for elemental and chemical information, enabling unique scientific outcomes. [ABSTRACT FROM AUTHOR]

The X‐ray fluorescence microscopy (XFM) beamline is an in‐vacuum undulator‐based X‐ray fluorescence (XRF) microprobe beamline at the 3 GeV Australian Synchrotron. The beamline delivers hard X‐rays in the 4–27 keV energy range, permitting K emission to Cd and L and M emission for all other heavier elements. With a practical low‐energy detection cut‐off of approximately 1.5 keV, low‐Z detection is constrained to Si, with Al detectable under favourable circumstances. The beamline has two scanning stations: a Kirkpatrick–Baez mirror microprobe, which produces a focal spot of 2 µm × 2 µm FWHM, and a large‐area scanning 'milliprobe', which has the beam size defined by slits. Energy‐dispersive detector systems include the Maia 384, Vortex‐EM and Vortex‐ME3 for XRF measurement, and the EIGER2 X 1 Mpixel array detector for scanning X‐ray diffraction microscopy measurements. The beamline uses event‐mode data acquisition that eliminates detector system time overheads, and motion control overheads are significantly reduced through the application of an efficient raster scanning algorithm. The minimal overheads, in conjunction with short dwell times per pixel, have allowed XFM to establish techniques such as full spectroscopic XANES fluorescence imaging, XRF tomography, fly scanning ptychography and high‐definition XRF imaging over large areas. XFM provides diverse analysis capabilities in the fields of medicine, biology, geology, materials science and cultural heritage. This paper discusses the beamline status, scientific showcases and future upgrades. [ABSTRACT FROM AUTHOR]

Owing to its extreme sensitivity, quantitative mapping of elemental distributions via X-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. [ABSTRACT FROM AUTHOR]

Highlights • A novel method using synchrotron-based micro X-ray fluorescence. • Provides laterally-resolved, multi-element, kinetic, in vivo analyses of plants. • Allows a wide range of studies for the examination of plant responses. Abstract The distribution and concentration of nutrients and contaminants affect almost every metabolic process in plants but analytical limitations have hindered the determination of microscopic changes over time within living plant tissues. We developed a novel method using synchrotron-based micro X-ray fluorescence (μ-XRF) that, for the first time, allows quantification of the spatial and temporal changes of multiple elements in the same area of living leaves. The utility of this approach was tested by examining changes over 48 h in unifoliate leaves of 7-d-old cowpea (Vigna unguiculata) plants simultaneously at 0.2 and 30 μM Mn in nutrient solution, with 30 μM Mn known to be toxic to cowpea and cause the formation of Mn-dense lesions. The fast X-ray fluorescence detector system reduced dwell on living leaf samples. This produced no evidence of tissue damage through repeated μ-XRF scanning, thereby overcoming previously noted experimental artifacts. This permitted, for the first time, visual and quantitative assessments of spatial and temporal changes in nutrient concentrations. By focusing on changes in Mn status, this study illustrated extension of two-dimensional μ-XRF scans to a three-dimensional geometry of Mn kinetics in the same area of leaves. The multi-element potential of this method was exemplified through the measurement of distributions and concentrations of K, Ca, Fe, Cu, and Zn within living plant leaves. This novel method and accompanying information on changes in Mn distribution showed the potential for microscopic, time-resolved, in vivo examination of changes in elemental distribution. We consider that this method will be of benefit for a wide range of studies, including functional characterization of molecular biology, examining changes in the distribution of nutrients, and understanding the movement and toxicity of contaminants. [ABSTRACT FROM AUTHOR]