Correlative 3D x-ray fluorescence and ptychographic tomography of frozen-hydrated green algae

Deng, Junjing; Lo, Yuan Hung; Gallagher-Jones, Marcus; Chen, Si; Pryor, Alan; Jin, Qiaoling; Hong, Young Pyo; Nashed, Youssef S. G.; Vogt, Stefan; Miao, Jianwei; Jacobsen, Chris

doi:10.1126/sciadv.aau4548

Cited by 93 publications

(86 citation statements)

References 70 publications

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“…However, these methods limit sample size well below the typical grain size in kesterite. Compared to other synchrotron techniques, RXPT probes a larger scale than XRD but provides real space information, and has superior resolution and accuracy for quantification than 3D fluorescence even in its most advanced correlative version [48]. Elemental sensitivity, which is the main advantage of fluorescence, is obtained in this case for the elements of interest at the cost of a change of energy and additional scans.…”

Section: E Outlookmentioning

confidence: 99%

Resonant x-ray ptychographic nanotomography of kesterite solar cells

Fevola

Jørgensen

Verezhak

et al. 2020

Phys. Rev. Research

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The Cu 2 ZnSnS 4 kesterite is currently among the most promising inorganic, nontoxic, earth-abundant materials for a new generation of solar cells. Interfacial defects and secondary phases present in the kesterite active layer are, however, detrimental to the performance of the device. They are typically probed with techniques that are destructive or limited to the surface, and x-ray diffraction cannot reliably distinguish small amounts of zinc sulfide or copper tin sulfide from kesterite. Conversely, resonant ptychographic tomography, which relies on electron density contrast, overcomes these limitations. Here, we demonstrate how this technique can enable localization and quantification of secondary phases, along with measurements of adherence at the interfacial layers, on complete and functioning devices. In our experiment, we utilize an x-ray energy value far from absorption edges as well as three single energies corresponding to the absorption edges of Cu, Zn, and Sn, to gain elemental sensitivity to these elements and enhance contrast between phases with similar electron density. As a result, we image and identify in the active layer grains of a secondary phase, namely, zinc sulfide, which is not easily discriminated by other standard characterization techniques. In addition, we are able to observe Cu diffused from the active layer into the CdS buffer layer as well as Cu in the form of copper sulfide at their interface. Other relevant morphological features are best resolved off-resonance at the optimal energy for the synchrotron beamline with ∼20 nm resolution.

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Section: E Outlookmentioning

confidence: 99%

Resonant x-ray ptychographic nanotomography of kesterite solar cells

Fevola

Jørgensen

Verezhak

et al. 2020

Phys. Rev. Research

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“…In particular, in combination with high-resolution X-ray microscopy, 3D nano-imaging is important for many applications in biology and materials science [1]. Two-dimensional sub-50 nm spatial resolution can be commonly reached in X-ray microscopy [2][3][4][5][6][7] for sample thickness of up to tens of microns. However, this high 2D resolution can be translated to comparable 3D resolution only if a sufficient number of measured angular 2D projections of the sample provide mutually consistent information.…”

Section: Introductionmentioning

confidence: 99%

Alignment methods for nanotomography with deep subpixel accuracy

et al. 2019

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As the resolution of X-ray tomography improves, the limited long-term stability and accuracy of nanoimaging tools does not allow computing artifact-free three-dimensional (3D) reconstructions without an additional step of numerical alignment of the measured projections. However, the common iterative alignment methods are significantly more computationally demanding than a simple tomographic reconstruction of the acquired volume. Here, we address this issue and present an alignment toolkit, which exploits methods with deep-subpixel accuracy combined with a multi-resolution scheme. This leads to robust and accurate alignment with significantly reduced computational and memory requirements. The performance of the presented methods is demonstrated on simulated and measured datasets for tomography and also laminography acquisition geometries. A GPU accelerated implementation of our alignment framework is publicly available.

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“…Both techniques were further extended by in-vacuum cryogenic sample cooling, providing simultaneously qualitative distributions of light elements within a cell and its morphology at remarkable sensitivity and sub-50-nm spatial resolution 27 . Further developments involved the implementation of a continuous-motion scanning scheme [28][29][30][31] and a sample rotation which enabled simultaneous acquisition of a tomographic dataset in both modalities with reduced time overhead 32 . Yet, these demonstrated solutions have until now featured an accessible field of view limited to the imaging of single cells.…”

mentioning

confidence: 99%

Multimodal X-ray imaging of nanocontainer-treated macrophages and calcium distribution in the perilacunar bone matrix

Stachnik

Warmer

Mohácsi

et al. 2020

Sci Rep

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Studies of biological systems typically require the application of several complementary methods able to yield statistically-relevant results at a unique level of sensitivity. Combined X-ray fluorescence and ptychography offer excellent elemental and structural imaging contrasts at the nanoscale. They enable a robust correlation of elemental distributions with respect to the cellular morphology. Here we extend the applicability of the two modalities to higher X-ray excitation energies, permitting iron mapping. Using a long-range scanning setup, we applied the method to two vital biomedical cases. We quantified the iron distributions in a population of macrophages treated with Mycobacteriumtuberculosis-targeting iron-oxide nanocontainers. Our work allowed to visualize the internalization of the nanocontainer agglomerates in the cytosol. From the iron areal mass maps, we obtained a distribution of antibiotic load per agglomerate and an average areal concentration of nanocontainers in the agglomerates. In the second application we mapped the calcium content in a human bone matrix in close proximity to osteocyte lacunae (perilacunar matrix). A concurrently acquired ptychographic image was used to remove the mass-thickness effect from the raw calcium map. The resulting ptychographyenhanced calcium distribution allowed then to observe a locally lower degree of mineralization of the perilacunar matrix. Metal ions play an important role in the vital functions of living organisms. They are present in various biological systems in a vast range of concentrations as structural, electrolyte (minor), and trace elements. From being major tissue components (e.g. Ca in bones) to constituents of essential biological molecules (e.g. Fe in hemoglobin), metals take part in the majority of extra-and intracellular processes. In particular, first-row transition metals (Mn, Fe, Cu, Ni, Zn)-despite their minute concentrations-are involved in sub-cellular processes. Moreover, their abnormal accumulation in human brain was correlated with mechanisms leading to neurodegeneration diseases such as Parkinson's and Alzheimer's 1-3. Metal compounds have also been utilized as novel drug delivery systems by addressing metabolic properties of bacterial agents 4,5 or as more efficient medical imaging markers tracking tissue of interest 6-8. In all aforementioned cases, studies of metal contributions require knowledge of their quantitative spatial distributions with respect to the sub-cellular structure. Electron-probe Energy-Dispersive Spectroscopy offers elemental mapping at nanometer-range spatial resolutions and high excitation efficiency in the low-Z-element

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Correlative 3D x-ray fluorescence and ptychographic tomography of frozen-hydrated green algae

Abstract: X-ray ptychography and fluorescence imaging reveal 3D elemental composition and ultrastructure in frozen-hydrated green algae.

Cited by 93 publications

References 70 publications

Resonant x-ray ptychographic nanotomography of kesterite solar cells

Resonant x-ray ptychographic nanotomography of kesterite solar cells

Alignment methods for nanotomography with deep subpixel accuracy

Multimodal X-ray imaging of nanocontainer-treated macrophages and calcium distribution in the perilacunar bone matrix

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