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X-ray ptychographic tomography is a nondestructive method for three dimensional (3D) imaging with nanometer-sized resolvable features. The size of the volume that can be imaged is almost arbitrary, limited only by the penetration depth and the available scanning time. Here we present a method that rapidly accelerates the imaging operation over a given volume through acquiring a limited set of data via large angular reduction and compensating for the resulting ill-posedness through deeply learned priors. The proposed 3D reconstruction method “RAPID” relies initially on a subset of the object measured with the nominal number of required illumination angles and treats the reconstructions from the conventional two-step approach as ground truth. It is then trained to reproduce equal fidelity from much fewer angles. After training, it performs with similar fidelity on the hitherto unexamined portions of the object, previously not shown during training, with a limited set of acquisitions. In our experimental demonstration, the nominal number of angles was 349 and the reduced number of angles was 21, resulting in a $$\times 140$$ × 140 aggregate speedup over a volume of $$4.48\times 93.18\times 3.92\, \upmu \text {m}^3$$ 4.48 × 93.18 × 3.92 μ m 3 and with $$(14\,\text {nm})^3$$ ( 14 nm ) 3 feature size, i.e. $$\sim 10^8$$ ∼ 10 8 voxels. RAPID’s key distinguishing feature over earlier attempts is the incorporation of atrous spatial pyramid pooling modules into the deep neural network framework in an anisotropic way. We found that adjusting the atrous rate improves reconstruction fidelity because it expands the convolutional kernels’ range to match the physics of multi-slice ptychography without significantly increasing the number of parameters.

Broadband X-ray ptychography is developed based on the multi-wavelength reconstruction method and is systematically demonstrated in both simulation and experiment. In addition, a combined reconstruction approach is proposed to further increase the ptychographic imaging rate., Ptychography is a rapidly developing scanning microscopy which is able to view the internal structures of samples at a high resolution beyond the illumination size. The achieved spatial resolution is theoretically dose-limited. A broadband source can provide much higher flux compared with a monochromatic source; however, it conflicts with the necessary coherence requirements of this coherent diffraction imaging technique. In this paper, a multi-wavelength reconstruction algorithm has been developed to deal with the broad bandwidth in ptychography. Compared with the latest development of mixed-state reconstruction approach, this multi-wavelength approach is more accurate in the physical model, and also considers the spot size variation as a function of energy due to the chromatic focusing optics. Therefore, this method has been proved in both simulation and experiment to significantly improve the reconstruction when the source bandwidth, illumination size and scan step size increase. It is worth mentioning that the accurate and detailed information of the energy spectrum for the incident beam is not required in advance for the proposed method. Further, we combine multi-wavelength and mixed-state approaches to jointly solve temporal and spatial partial coherence in ptychography so that it can handle various disadvantageous experimental effects. The significant relaxation in coherence requirements by our approaches allows the use of high-flux broadband X-ray sources for high-efficient and high-resolution ptychographic imaging.

Single-crystal cathode materials for lithium-ion batteries have attracted increasing interest in providing greater capacity retention than their polycrystalline counterparts. However, after being cycled at high voltages, these single-crystal materials exhibit severe structural instability and capacity fade. Understanding how the surface structural changes determine the performance degradation over cycling is crucial, but remains elusive. Here, we investigate the correlation of the surface structure, internal strain, and capacity deterioration by using operando X-ray spectroscopy imaging and nano-tomography. We directly observe a close correlation between surface chemistry and phase distribution from homogeneity to heterogeneity, which induces heterogeneous internal strain within the particle and the resulting structural/performance degradation during cycling. We also discover that surface chemistry can significantly enhance the cyclic performance. Our modified process effectively regulates the performance fade issue of single-crystal cathode and provides new insights for improved design of high-capacity battery materials., Performance improvement of cathode materials represents one of the most critical technological challenges for lithium-ion batteries. Here the authors show a close correlation between surface chemistry and phase distribution from homogeneity to heterogeneity, and its impact on the cyclic performance.

In this article, a systematic framework for designing the control for fine positioning (scanning) stages of X-ray microscopes is presented. This framework facilitates designs that simultaneously achieve specifications on positioning resolution and tracking bandwidth while guaranteeing robustness of the closed loop device to unmodeled uncertainties. We use robust optimal control techniques for modeling, quantifying design objectives and system-specific challenges, and designing the control laws. The control designs were implemented on a three degree of freedom piezoactuated flexure stages dedicated for fine positioning of X-ray optics. Experimental results demonstrate significant improvements in positioning performance of 134%, 150%, and 132% in tracking bandwidths along the lateral ( X ), vertical ( Y ), and beam ( Z ) directions, respectively, when compared to proportional–integral–derivative controller designs. This was achieved while keeping similar or better positioning resolution and robustness measures. Fast scanning for X-ray imaging was demonstrated in both the step scan and flyscan modes, where bandwidth was improved by over 450 times with flyscan compared to the step scan.

Summary Halide perovskites have emerged as a new class of inorganic-based soft semiconductors that are revolutionizing the field of (opto)electronics. However, the fundamental structures and properties or functions of halide perovskites are still far from being fully understood, calling for the need of precise, quantitative, and high-spatiotemporal-resolution characterizations. As one of the most powerful multimodal characterization techniques, synchrotron radiation offers high-brilliance X-ray beams for high-resolution observations across multiple length scales obtained within extremely short time spans. Performing a brief review of synchrotron-based perovskite studies in the literature, here we provide perspectives on the use of synchrotron characterizations not only for studying multiple-length-scale structures and dynamic variations of perovskite materials, but also for in situ monitoring of device functions and dynamics. The goal of this perspective is to present many capabilities and evoke emerging opportunities of synchrotron characterizations for providing decisive answers to the outstanding science problems in the perovskite field, pushing forward the technological development.

X-ray ptychography imaging at synchrotron facilities like the Advanced Photon Source (APS) involves controlling instrument hardwares to collect a set of diffraction patterns from overlapping coherent illumination spots on extended samples, managing data storage, reconstructing ptychographic images from acquired diffraction patterns, and providing the visualization of results and feedback. In addition to the complicated workflow, ptychography instrument could produce up to several TB's of data per second that is needed to be processed in real time. This brings up the need to develop a high performance, robust and user friendly processing software package for ptychographic data analysis. In this paper we present a software framework which provides functionality of visualization, work flow control, and data reconstruction. To accelerate the computation and large datasets process, the data reconstruction part is implemented with three algorithms, ePIE, DM and LSQML using CUDA-C on GPU., Comment: X-Ray Nanoimaging: Instruments and Methods V

Simple Summary Metal-oxide nanomaterials enter cancer and normal cells even when not specifically targeted, and often interact with specific cellular structures and biological molecules solely due to their innate physical-chemical properties. This raises concerns for the use of nanoparticles, which can be alleviated only with rigorous studies of nanoparticle–cell interactions, studies independent of post-interaction labeling of nanomaterials. X-ray fluorescence microscopy is an imaging technique that quantifies and maps all chemical elements from the periodic table solely based on their native fluorescence excited by the incoming X-ray. We used two different instruments to interrogate the same sample in 3D at two different resolutions and determine heterogeneity of cell-to-cell interactions with nanomaterials, as well as subcellular nanoparticle distribution. This is the first example of multi-scale 3D X-ray fluorescence imaging. This work begins a new era of study on how nanoparticle-based therapies can be developed to be more predictable and safer for use. Abstract Research in cancer nanotechnology is entering its third decade, and the need to study interactions between nanomaterials and cells remains urgent. Heterogeneity of nanoparticle uptake by different cells and subcellular compartments represent the greatest obstacles to a full understanding of the entire spectrum of nanomaterials’ effects. In this work, we used flow cytometry to evaluate changes in cell cycle associated with non-targeted nanocomposite uptake by individual cells and cell populations. Analogous single cell and cell population changes in nanocomposite uptake were explored by X-ray fluorescence microscopy (XFM). Very few nanoparticles are visible by optical imaging without labeling, but labeling increases nanoparticle complexity and the risk of modified cellular uptake. XFM can be used to evaluate heterogeneity of nanocomposite uptake by directly imaging the metal atoms present in the metal-oxide nanocomposites under investigation. While XFM mapping has been performed iteratively in 2D with the same sample at different resolutions, this study is the first example of serial tomographic imaging at two different resolutions. A cluster of cells exposed to non-targeted nanocomposites was imaged with a micron-sized beam in 3D. Next, the sample was sectioned for immunohistochemistry as well as a high resolution “zoomed in” X-ray fluorescence (XRF) tomography with 80 nm beam spot size. Multiscale XRF tomography will revolutionize our ability to explore cell-to-cell differences in nanomaterial uptake.

Examining chemical and structural characteristics of micro-features in complex tissue matrices is essential for understanding biological systems. Advances in multimodal chemical and structural imaging using synchrotron radiation have overcome many issues in correlative imaging, enabling the characterization of distinct microfeatures at nanoscale resolution in ex vivo tissues. We present a nanoscale imaging method that pairs X-ray ptychography and X-ray fluorescence microscopy (XFM) to simultaneously examine structural features and quantify elemental content of microfeatures in complex ex vivo tissues. We examined the neuropathological microfeatures Lewy bodies, aggregations of superoxide dismutase 1 (SOD1) and neuromelanin in human post-mortem Parkinson's disease tissue. Although biometals play essential roles in normal neuronal biochemistry, their dyshomeostasis is implicated in Parkinson's disease aetiology. Here we show that Lewy bodies and SOD1 aggregates have distinct elemental fingerprints yet are similar in structure, whilst neuromelanin exhibits different elemental composition and a distinct, disordered structure. The unique approach we describe is applicable to the structural and chemical characterization of a wide range of complex biological tissues at previously unprecedented levels of detail., Structural and chemical characterisation of microfeatures in unadulterated Parkinson's disease brain tissue using synchrotron nanoscale XFM and ptychography.

Scanning of lightweight circular diffractive optics, separate from central stops and apertures, is emerging as an approach to exploit advances in synchrotron x-ray sources. We consider the effects in a scanning microscope of offsets between the optic and its central stop and find that scan ranges of up to about half the diameter of the optic are possible with only about a 10% increase in the focal spot width. For large scanning ranges, we present criteria for the working distance between the last aperture and the specimen to be imaged.

The lack of precision therapies combined with limited therapeutic access to intracranial tumor sites due to the presence of the blood-brain/blood-tumor barriers have contributed to glioblastoma (GBM) being one of the most difficult cancers to effectively treat. We have developed a novel precision medicine approach for GBM treatment that involves the use of brain-penetrant RNA interference (RNAi)-based spherical nucleic acids (SNAs). In this study, we used SNAs consisting of gold nanoparticle cores covalently conjugated with radially oriented and densely packed siRNA oligonucleotides. Based upon previous preclinical evaluation, we conducted toxicology and toxicokinetic studies in non-human primates, and a single-arm, open-label phase 0 first-in-human trial (NCT03020017) to determine safety, pharmacokinetics, intratumoral accumulation and gene-suppressive activity of systemically administered SNAs carrying siRNA specific for the GBM oncogene Bcl2Like12 (Bcl2L12). Patients with recurrent GBM were treated with intravenous administration of siBcl2L12-SNAs (drug moniker: NU-0129), at a dose corresponding to 1/50th of the no-observed-adverse-event-level (NOAEL), followed by tumor resection. Safety assessment revealed no significant treatment-related toxicities. Inductively coupled plasma mass spectrometry, X-ray fluorescence microscopy, and silver staining of resected GBM tissue demonstrated that intravenously administered SNAs reached patient tumors, with gold (Au) enrichment observed in the tumor-associated endothelium, macrophages and tumor cells. NU-0129 uptake into glioma cells correlated with significant reduction in tumor-associated Bcl2L12 protein expression, as indicated by comparison of NU-0129-treated recurrent vs. matched primary (i.e., untreated) tumor. Our results establish SNA nanoconjugates as a brain-penetrant precision medicine approach for the systemic treatment of GBM.

As a coherent diffraction imaging technique, ptychography provides high-spatial resolution beyond Rayleigh’s criterion of the focusing optics, but it is also sensitively affected by the decoherence coming from the spatial and temporal variations in the experiment. Here we show that high-speed ptychographic data acquisition with short exposure can effectively reduce the impact from experimental variations. To reach a cumulative dose required for a given resolution, we further demonstrate that a continuous multi-pass scan via high-speed ptychography can achieve high-resolution imaging. This low-dose scan strategy is shown to be more dose-efficient, and has potential for radiation-sensitive sample studies and time-resolved imaging.

Additional file 1. Additional tables and figures.

Supplemental Document

We describe and demonstrate an optimization-based X-ray image reconstruction framework called Adorym. Our framework provides a generic forward model, allowing one code framework to be used for a wide range of imaging methods ranging from near-field holography to fly-scan ptychographic tomography. By using automatic differentiation for optimization, Adorym has the flexibility to refine experimental parameters including probe positions, multiple hologram alignment, and object tilts. It is written with strong support for parallel processing, allowing large datasets to be processed on high-performance computing systems. We demonstrate its use on several experimental datasets to show improved image quality through parameter refinement. (C) 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

Background Neuroblastoma is the most common extracranial solid malignancy in childhood which, despite the current progress in radiotherapy and chemotherapy protocols, still has a high mortality rate in high risk tumors. Nanomedicine offers exciting and unexploited opportunities to overcome the shortcomings of conventional medicine. The photocatalytic properties of Fe3O4 core-TiO2 shell nanocomposites and their potential for cell specific targeting suggest that nanoconstructs produced using Fe3O4 core-TiO2 shell nanocomposites could be used to enhance radiation effects in neuroblastoma. In this study, we evaluated bare, metaiodobenzylguanidine (MIBG) and 3,4-Dihydroxyphenylacetic acid (DOPAC) coated Fe3O4@TiO2 as potential radiosensitizers for neuroblastoma in vitro. Results The uptake of bare and MIBG coated nanocomposites modestly sensitized neuroblastoma cells to ionizing radiation. Conversely, cells exposed to DOPAC coated nanocomposites exhibited a five-fold enhanced sensitivity to radiation, increased numbers of radiation induced DNA double-strand breaks, and apoptotic cell death. The addition of a peptide mimic of the epidermal growth factor (EGF) to nanoconjugates coated with MIBG altered their intracellular distribution. Cryo X-ray fluorescence microscopy tomography of frozen hydrated cells treated with these nanoconjugates revealed cytoplasmic as well as nuclear distribution of the nanoconstructs. Conclusions The intracellular distribution pattern of different nanoconjugates used in this study was different for different nanoconjugate surface molecules. Cells exposed to DOPAC covered nanoconjugates showed the smallest nanoconjugate uptake, with the most prominent pattern of large intracellular aggregates. Interestingly, cells treated with this nanoconjugate also showed the most pronounced radiosensitization effect in combination with the external beam x-ray irradiation. Further studies are necessary to evaluate mechanistic basis for this increased radiosensitization effect. Preliminary studies with the nanoparticles carrying an EGF mimicking peptide showed that this approach to targeting could perhaps be combined with a different approach to radiosensitization – use of nanoconjugates in combination with the radioactive iodine. Much additional work will be necessary in order to evaluate possible benefits of targeted nanoconjugates carrying radionuclides. Graphic abstract Supplementary Information The online version contains supplementary material available at 10.1186/s12645-021-00081-z.

X-ray ptychography is a rapidly developing coherent diffraction imaging technique that provides nanoscale resolution on extended field-of-view. However, the requirement of coherence and the scanning mechanism limit the throughput of ptychographic imaging. In this paper, we propose X-ray ptychography using multiple illuminations instead of single illumination in conventional ptychography. Multiple locations of the sample are simultaneously imaged by spatially separated X-ray beams, therefore, the obtained field-of-view in one scan can be enlarged by a factor equal to the number of illuminations. We have demonstrated this technique experimentally using two X-ray beams focused by a house-made Fresnel zone plate array. Two areas of the object and corresponding double illuminations were successfully reconstructed from diffraction patterns acquired in one scan, with image quality similar with those obtained by conventional single-beam ptychography in sequence. Multi-beam ptychography approach increases the imaging speed, providing an efficient way for high-resolution imaging of large extended specimens.

The development of commercially friendly and stable catalysts for oxygen reduction reaction (ORR) is critical for many energy conversion systems such as fuel cells and metal-air batteries. Many Co-based perovskite oxides such as LaCoO3 have been discovered as the stable and active ORR catalysts, which can be good candidates to replace platinum (Pt). Although researchers have tried substituting various transition metals into the Co-based perovskite catalysts to improve the ORR performance, the influence of substitution on the ORR mechanism is rarely studied. In this paper, we explore the evolution of ORR mechanism after substituting Fe into LaCoO3, using the combination of X-ray photoelectron spectroscopy, high-resolution X-ray microscopy, X-ray diffraction, surface-sensitive soft X-ray absorption spectroscopy characterization, and electrochemical tests. We observed enhanced catalytic activities and increased electron transfer numbers during the ORR in Co-rich perovskite, which are attributed to the optimized eg filling numbers and the stronger hybridization of transition metal 3d and oxygen 2p bands. The discoveries in this paper provide deep insights into the ORR catalysis mechanism on metal oxides and new guidelines for the design of Pt-free ORR catalysts.

X-ray ptychography, a powerful scanning lensless imaging technique, has become attractive for nondestructively imaging internal structures at nanoscale. Stage positioning overhead in conventional step-scan ptychography is one of the limiting factors on the imaging throughput. In this work, we demonstrate the use of advanced fly scan ptychography to achieve high-resolution ptychograms of modern integrated circuits on a large field-of-view at millimeter scale. By completely removing stage overheads between scan points, the imaging time for millimeter-size sample can be significantly reduced. Furthermore, we implement the orthogonal probe relaxation technique to overcome the variation of illumination across the large scan area as well as local vibrations. The capability of x-ray ptychography shown here is broadly applicable for various studies, which requires both high spatial resolution and large scan area.

X-ray ptychography is being utilized for a wide range of imaging experiments with a resolution beyond the limit of the X-ray optics used. Here, strategies for data sampling and for increasing imaging throughput when the specimen is at the focus of an X-ray beam are discussed, and the tradeoffs between large and small illumination spots are examined., X-ray ptychography is being utilized for a wide range of imaging experiments with a resolution beyond the limit of the X-ray optics used. Introducing a parameter for the ptychographic resolution gain G p (the ratio of the beam size over the achieved pixel size in the reconstructed image), strategies for data sampling and for increasing imaging throughput when the specimen is at the focus of an X-ray beam are considered. The tradeoffs between large and small illumination spots are examined.

State-of-the-art halide perovskite solar cells have bandgaps larger than 1.45 eV, which restricts their potential for realizing the Shockley-Queisser limit. Previous search for low-bandgap (1.2 to 1.4 eV) halide perovskites has resulted in several candidates, but all are hybrid organic-inorganic compositions, raising potential concern regarding device stability. Here we show the promise of an inorganic low-bandgap (1.38 eV) CsPb0.6Sn0.4I3 perovskite stabilized via interface functionalization. Device efficiency up to 13.37% is demonstrated. The device shows high operational stability under one-sun-intensity illumination, with T80 and T70 lifetimes of 653 h and 1045 h, respectively (T80 and T70 represent efficiency decays to 80% and 70% of the initial value, respectively), and long-term shelf stability under nitrogen atmosphere. Controlled exposure of the device to ambient atmosphere during a long-term (1000 h) test does not degrade the efficiency. These findings point to a promising direction for achieving low-bandgap perovskite solar cells with high stability., Current research focus on the perovskites solar cells (PSCs) is mainly limited to the lead-based ones with bandgaps above 1.5 eV. Here Hu et al. report efficient and stable inorganic tin-containing PSCs, opening doors to exploring abundant perovskite materials with bandgaps lower than 1.4 eV.

Synchrotron radiation light source facilities are leading the way to ultrahigh resolution X-ray imaging. High resolution imaging is essential to understanding the fundamental structure and interaction of materials at the smallest length scale possible. Diffraction based methods achieve nanoscale imaging by replacing traditional objective lenses by pixelated area detectors and computational image reconstruction. Among these methods, ptychography is quickly becoming the standard for sub-30 nanometer imaging of extended samples, but at the expense of increasingly high data rates and volumes. This paper presents a new distributed algorithm for solving the ptychographic image reconstruction problem based on automatic differentiation. Input datasets are subdivided between multiple graphics processing units (GPUs); each subset of the problem is then solved either entirely independent of other subsets (asynchronously) or through sharing gradient information with other GPUs (synchronously). The algorithm was evaluated on simulated and real data acquired at the Advanced Photon Source, scaling up to 192 GPUs. The synchronous variant of our method outperformed an existing multi-GPU implementation in terms of accuracy while running at a comparable execution time.

Achieving high spatial resolution is the goal of many imaging systems. Designing a high-resolution lens with diffraction-limited performance over a large field of view remains a difficult task in imaging system design. On the other hand, creating a complex speckle pattern with wavelength-limited spatial features is effortless and can be implemented via a simple random diffuser. With this observation and inspired by the concept of near-field ptychography, we report a new imaging modality, termed near-field Fourier ptychography, for tackling high-resolution imaging challenges in both microscopic and macroscopic imaging settings. The meaning of 'near-field' is referred to placing the object at a short defocus distance with a large Fresnel number. In our implementations, we project a speckle pattern with fine spatial features on the object instead of directly resolving the spatial features via a high-resolution lens. We then translate the object (or speckle) to different positions and acquire the corresponding images using a low-resolution lens. A ptychographic phase retrieval process is used to recover the complex object, the unknown speckle pattern, and the coherent transfer function at the same time. In a microscopic imaging setup, we use a 0.12 numerical aperture (NA) lens to achieve a NA of 0.85 in the reconstruction process. In a macroscale photographic imaging setup, we achieve ~7-fold resolution gain using a photographic lens. The final achievable resolution is not determined by the collection optics. Instead, it is determined by the feature size of the speckle pattern. The reported imaging modality can be employed in light, coherent X-ray, and transmission electron imaging systems to increase resolution and provide quantitative absorption and phase contrast of the object., Comment: 15 pages, 14 figures

X-ray ptychography and fluorescence imaging reveal 3D elemental composition and ultrastructure in frozen-hydrated green algae., Accurate knowledge of elemental distributions within biological organisms is critical for understanding their cellular roles. The ability to couple this knowledge with overall cellular architecture in three dimensions (3D) deepens our understanding of cellular chemistry. Using a whole, frozen-hydrated Chlamydomonas reinhardtii cell as an example, we report the development of 3D correlative microscopy through a combination of simultaneous cryogenic x-ray ptychography and x-ray fluorescence microscopy. By taking advantage of a recently developed tomographic reconstruction algorithm, termed GENeralized Fourier Iterative REconstruction (GENFIRE), we produce high-quality 3D maps of the unlabeled alga’s cellular ultrastructure and elemental distributions within the cell. We demonstrate GENFIRE’s ability to outperform conventional tomography algorithms and to further improve the reconstruction quality by refining the experimentally intended tomographic angles. As this method continues to advance with brighter coherent light sources and more efficient data handling, we expect correlative 3D x-ray fluorescence and ptychographic tomography to be a powerful tool for probing a wide range of frozen-hydrated biological specimens, ranging from small prokaryotes such as bacteria, algae, and parasites to large eukaryotes such as mammalian cells, with applications that include understanding cellular responses to environmental stimuli and cell-to-cell interactions.

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.

X-Ray Microscopy 2018, Saskatoon, Canada, 19 Aug 2018 - 24 Aug 2018; Microscopy and microanalysis 24(S2), 434 - 435 (2018). doi:10.1017/S1431927618014423, Published by Cambridge University Press

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.

Modern integrated circuits (ICs) employ a myriad of materials organized at nanoscale dimensions, and certain critical tolerances must be met for them to function. To understand departures from intended functionality, it is essential to examine ICs as manufactured so as to adjust design rules, ideally in a non-destructive way so that imaged structures can be correlated with electrical performance. Electron microscopes can do this on thin regions, or on exposed surfaces, but the required processing alters or even destroys functionality. Microscopy with multi-keV x-rays provides an alternative approach with greater penetration, but the spatial resolution of x-ray imaging lenses has not allowed one to see the required detail in the latest generation of ICs. X-ray ptychography provides a way to obtain images of ICs without lens-imposed resolution limits, with past work delivering 20–40 nm resolution on thinned ICs. We describe a simple model for estimating the required exposure, and use it to estimate the future potential for this technique. Here we show for the first time that this approach can be used to image circuit detail through an unprocessed 300 μm thick silicon wafer, with sub-20 nm detail clearly resolved after mechanical polishing to 240 μm thickness was used to eliminate image contrast caused by Si wafer surface scratches. By using continuous x-ray scanning, massively parallel computation, and a new generation of synchrotron light sources, this should enable entire non-etched ICs to be imaged to 10 nm resolution or better while maintaining their ability to function in electrical tests.

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.

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.

Ptychography has emerged as a nondestructive tool to quantitatively study extended samples at a high spatial resolution. In this manuscript, we report on recent developments from our team. We have combined cryo ptychography and fluorescence microscopy to provide simultaneous views of ultrastructure and elemental composition, we have developed multi-GPU parallel computation to speed up ptychographic reconstructions, and we have implemented fly-scan ptychography to allow for faster data acquisition. We conclude with a discussion of future challenges in high-resolution 3D ptychography.

Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object’s complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous “fly-scan” mode for ptychographic data collection in which the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.