Nanoscale Imaging Techniques Research Articles

Protein fibril aggregation on red blood cells: a potential biomarker to distinguish neurodegenerative diseases from healthy aging.

Neurodegenerative diseases like Alzheimer's disease are characterized by the accumulation of misfolded proteins into fibrils in the brain. Atomic force microscopy is a nanoscale imaging technique that can be used to resolve and quantify protein aggregates from oligomers to fibrils. Recently, we characterized protein fibrillar aggregates adsorbed on the surface of red blood cells with atomic force microscopy from patients with neurocognitive disorders, suggesting a novel Alzheimer's disease biomarker. However, the age association of fibril deposits on red blood cells has not yet been studied in detail in healthy adults. Here, we used atomic force microscopy to visualize and quantify fibril coverage on red blood cells in 50 healthy adults and 37 memory clinic patients. Fibrillar protein deposits sporadically appeared in healthy individuals but were much more prevalent in patients with neurodegenerative disease, especially those with Alzheimer's disease as confirmed by positive CSF amyloid beta 1-42/1-40 ratios. The prevalence of fibrils on the red blood cell surface did not significantly correlate with age in either healthy individuals or Alzheimer's disease patients. The overlap in fibril prevalence on red blood cells between Alzheimer's disease and amyloid-negative patients suggests that fibril deposition on red blood cells could occur in various neurodegenerative diseases. Quantifying red blood cell protein fibril morphology and prevalence on red blood cells could serve as a sensitive biomarker for neurodegeneration, distinguishing between healthy individuals and those with neurodegenerative diseases. Future studies that combine atomic force microscopy with immunofluorescence techniques in larger-scale studies could further identify the chemical nature of these fibrils, paving the way for a comprehensive, non-invasive biomarker platform for neurodegenerative diseases.

Harnessing Nanotechnology: Emerging Strategies for Multiple Myeloma Therapy.

Advances in nanotechnology have provided novel avenues for the diagnosis and treatment of multiple myeloma (MM), a hematological malignancy characterized by the clonal proliferation of plasma cells in the bone marrow. This review elucidates the potential of nanotechnology to revolutionize myeloma therapy, focusing on nanoparticle-based drug delivery systems, nanoscale imaging techniques, and nano-immunotherapy. Nanoparticle-based drug delivery systems offer enhanced drug targeting, reduced systemic toxicity, and improved therapeutic efficacy. We discuss the latest developments in nanocarriers, such as liposomes, polymeric nanoparticles, and inorganic nanoparticles, used for the delivery of chemotherapeutic agents, siRNA, and miRNA in MM treatment. We delve into nanoscale imaging techniques which provide spatial multi-omic data, offering a holistic view of the tumor microenvironment. This spatial resolution can help decipher the complex interplay between cancer cells and their surrounding environment, facilitating the development of highly targeted therapies. Lastly, we explore the burgeoning field of nano-immunotherapy, which employs nanoparticles to modulate the immune system for myeloma treatment. Specifically, we consider how nanoparticles can be used to deliver tumor antigens to antigen-presenting cells, thus enhancing the body's immune response against myeloma cells. In conclusion, nanotechnology holds great promise for improving the prognosis and quality of life of MM patients. However, several challenges remain, including the need for further preclinical and clinical trials to assess the safety and efficacy of these emerging strategies. Future research should also focus on developing personalized nanomedicine approaches, which could tailor treatments to individual patients based on their genetic and molecular profiles.

Generic arrays of surface-positioned and shallow-buried gold multi-shapes as reference samples to benchmark near-field microscopes. Part 1: Applications in s-SNOM depth imaging

A wide palette of nanoscale imaging techniques operating in the near-field regime has been reported to date, enabling an important number of scientific breakthroughs. While the tuning and benchmarking of near-field microscopes represent a very important step for optimizing the outputs of the imaging sessions, no generally acknowledged standards exist yet in terms of calibration of near-field microscopes, which would play an important role in fully exploiting the potential of these instruments. With this work, we aim to contribute to filling in this gap, by introducing a prototypical sample, that holds potential for becoming a benchmark with respect to comparing the performances of diverse near-field measurement techniques, including traditional, aperture based, scanning near field microscopy (SNOM), or apertureless variants, such as scattering-type scanning nearfield optical microscopy (s-SNOM). The proposed samples have been thoroughly simulated, and an easy fabrication procedure is presented and demonstrated. In this latter context, Au-SiO2 samples sharing different configurations, in terms of geometry, number and depth of contrast yielding layers, enabling both surface and sub-surface nanoscopy measurements, were designed and fabricated. We argue that the proposed prototypical samples can be highly useful for benchmarking the outputs of various near-field microscopy techniques, as they facilitate a broad range of tests, relevant for comparing the performances and accuracy of many diverse investigation methods. We also introduce a methodology for numerically simulating the samples and their near-field after illuminating them with light of different wavelengths, as well as their simple process flow. This methodology can considerably augment their future use as a prototypical sample for the evaluation and calibration of current and next generation near-field nanoscopy techniques. Experimental evidence on the usefulness of these samples as s-SNOM testing and benchmarking tools is provided in the context of differentiation of surface and sub-surface structures, and influence of tip-sample distance on attainable amplitude and phase signals. We consider these efforts to represent an important, required step, in advancing the near-field imaging field, with important potential to augment the outputs of current near-field imaging systems, and to facilitate the development and benchmarking of next generation of near-field instrumentation.

Electrodeposition Parameters Dramatically Influence the Morphology, Stability, and Performance of n‐Si/Pt Light‐Addressable Electrochemical Sensors

AbstractLight addressable electrochemical (LAE) sensors have seen great utility in the past several years because they enable multiple localized electrochemical measurements to be performed on a single macroscopic electrode, opening up applications in imaging, biosensing, surface patterning, and multiplexing. In this study, we investigated the effects of electrodeposition on the formation of LAE sensors formed between n‐Si and electrodeposited Pt. We prepared sensors by electrodepositing Pt onto freshly‐etched n‐Si under a variety of conditions, varying the Pt precursor concentration, electrodeposition time, supporting electrolyte, and potential waveform. We characterized the sensors using a combination of atomic force microscopy, electrochemical impedance spectroscopy, and cyclic voltammetry. This study shows that the electrodeposition parameters have a dramatic impact on the morphology of the electrodeposited surfaces, sensor stability, and sensitivity towards H2O2. Specifically, we observed that continuous Pt films prepared with a higher Pt precursor concentration were more stable and had better linearity, higher sensitivity, and broader dynamic range than those prepared with a lower Pt precursor concentration. The stability and increased H2O2 sensing performance correlate strongly with an increase in the Pt islands which make up the film. These data highlight that the morphology of the metal in semiconductor/metal junction LAE sensors has an impact on important performance metrics like stability and sensitivity. They also demonstrate the need for semiconductor/metal LAE sensors to be studied using micro‐ and nanoscale imaging techniques in order to more deeply understand their performance characteristics.

Role of Plasmonic Antenna in Hot Carrier-Driven Reactions on Bimetallic Nanostructures.

Noble metal nanostructures can efficiently harvest electromagnetic radiation, which, in turn, is used to generate localized surface plasmon resonances. Surface plasmons decay, producing hot carriers, that is, short-lived species that can trigger chemical reactions on metallic surfaces. However, noble metal nanostructures catalyze only a very small number of chemical reactions. This limitation can be overcome by coupling such nanostructures with catalytic-active metals. Although the role of such catalytically active metals in plasmon-driven catalysis is well-understood, the mechanistics of a noble metal antenna in such chemistry remains unclear. In this study, we utilize tip-enhanced Raman spectroscopy, an innovative nanoscale imaging technique, to investigate the rates and yields of plasmon-driven reactions on mono- and bimetallic gold- and silver-based nanostructures. We found that silver nanoplates (AgNPs) demonstrate a significantly higher yield of 4-nitrobenzenehtiol to p,p'-dimercaptoazobisbenzene (DMAB) reduction than gold nanoplates (AuNPs). We also observed substantially greater yields of DMAB on silver-platinum and silver-palladium nanoplates (Ag@PtNPs and Ag@PdNPs) compared to their gold analogues, Au@PtNPs and Au@PdNPs. Furthermore, Ag@PtNPs exhibited enhanced reactivity in 4-mercatophenylmethanol to 4-mercaptobenzoic acid oxidation compared to Au@PtNPs. These results showed that silver-based bimetallic nanostructures feature much greater reactivity compared to their gold-based analogues.

Advances of Nanotechnology in the Diagnosis and Treatment of Hepatocellular Carcinoma.

Nanotechnology has emerged as a promising technology in the field of hepatocellular carcinoma (HCC), specifically in the implementation of diagnosis and treatment strategies. Nanotechnology-based approaches, such as nanoparticle-based contrast agents and nanoscale imaging techniques, have shown great potential for enhancing the sensitivity and specificity of HCC detection. These approaches provide high-resolution imaging and allow for the detection of molecular markers and alterations in cellular morphology associated with HCC. In terms of treatment, nanotechnology has revolutionized HCC therapy by enabling targeted drug delivery, enhancing therapeutic efficacy, and minimizing off-target effects. Nanoparticle-based drug carriers can be functionalized with ligands specific to HCC cells, allowing for selective accumulation of therapeutic agents at the tumor site. Furthermore, nanotechnology can facilitate combination therapy by co-encapsulating multiple drugs within a single nanoparticle, allowing for synergistic effects and overcoming drug resistance. This review aims to provide an overview of recent advances in nanotechnology-based approaches for the diagnosis and treatment of HCC. Further research is needed to optimize the design and functionality of nanoparticles, improve their biocompatibility and stability, and evaluate their long-term safety and efficacy. Nonetheless, the integration of nanotechnology in HCC management holds great promise and may lead to improved patient outcomes in the future.

Taking Bragg Coherent Diffraction Imaging to Higher Energies at Fourth Generation Synchrotrons: Nanoscale Characterization

High-energy Bragg coherent diffraction imaging (BCDI) can enable three-dimensional imaging of atomic structure within individual crystallites in complex environments. Here, we show that sufficient coherent photon flux is available to extend the BCDI technique to higher energies to (1) obtain improved strain information and sensitivity at the nanoscale with higher order Bragg reflections, (2) exploit BCDI in embedded materials or complex operando environments, (3) reduce X-ray induced sample modification, and (4) minimize dynamical scattering effects. We demonstrate the nanoscale imaging technique on the same sub-micrometer sized crystal at 8.5, 19.9, and 33.4 keV by taking advantage of the brilliance and coherence of the fourth generation Extremely Brilliant Source of the ID01 beamline at ESRF - The European Synchrotron (Grenoble, France). The photon flux, data quality, resolution and strain information are compared. We also show the first coherent diffraction measurements performed at the ID31-EBS beamline at an energy of 41 keV.

Enhancing optical contrast of sub-wavelength optical image using graphene oxide coated probe in scanning thermal microscopy

Scanning thermal microscopy (SThM) is a powerful technique for obtaining local thermal information on a sample surface using a nano-fabricated thermometer probe. One important application of SThM is sub-wavelength optical imaging, which captures the photothermal response of the probe to light. In this nano-scale imaging technique, optical contrast and spatial resolution are critical performance indicators. The size and material of the thermometer probe play a significant role in determining image quality, as a smaller radius can improve spatial resolution but reduce optical contrast and sensitivity due to decreased light absorption. To address this challenge, we have developed an approach to improve the photothermal energy conversion efficiency by coating the probe with atomically thin graphene oxide (GO). Our results demonstrate that this GO coating can enhance optical sensitivity by more than 1.5 times with linear optical power dependency. Importantly, the coating significantly improves the root mean square contrast of the image by 2.56 times without compromising spatial resolution. Our study highlights the potential of GO-coated probes to improve the sensitivity and quality of SThM-based sub-wavelength optical imaging.

Direct Imaging of the Kinetic Crystallization Pathway: Simulation and Liquid-Phase Transmission Electron Microscopy Observations.

The crystallization of materials from a suspension determines the structure and function of the final product, and numerous pieces of evidence have pointed out that the classical crystallization pathway may not capture the whole picture of the crystallization pathways. However, visualizing the initial nucleation and further growth of a crystal at the nanoscale has been challenging due to the difficulties of imaging individual atoms or nanoparticles during the crystallization process in solution. Recent progress in nanoscale microscopy had tackled this problem by monitoring the dynamic structural evolution of crystallization in a liquid environment. In this review, we summarized several crystallization pathways captured by the liquid-phase transmission electron microscopy technique and compared the observations with computer simulation. Apart from the classical nucleation pathway, we highlight three nonclassical pathways that are both observed in experiments and computer simulations: formation of an amorphous cluster below the critical nucleus size, nucleation of the crystalline phase from an amorphous intermediate, and transition between multiple crystalline structures before achieving the final product. Among these pathways, we also highlight the similarities and differences between the experimental results of the crystallization of single nanocrystals from atoms and the assembly of a colloidal superlattice from a large number of colloidal nanoparticles. By comparing the experimental results with computer simulations, we point out the importance of theory and simulation in developing a mechanistic approach to facilitate the understanding of the crystallization pathway in experimental systems. We also discuss the challenges and future perspectives for investigating the crystallization pathways at the nanoscale with the development of in situ nanoscale imaging techniques and potential applications to the understanding of biomineralization and protein self-assembly.

Membrane Remodeling of Human-Engineered Cardiac Tissue by Chronic Electric Stimulation.

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show immature features, but these are improved by integration into 3D cardiac constructs. In addition, it has been demonstrated that physical manipulations such as electrical stimulation (ES) are highly effective in improving the maturation of human-engineered cardiac tissue (hECT) derived from hiPSC-CMs. Here, we continuously applied an ES in capacitive coupling configuration, which is below the pacing threshold, to millimeter-sized hECTs for 1-2 weeks. Meanwhile, the structural and functional developments of the hECTs were monitored and measured using an array of assays. Of particular note, a nanoscale imaging technique, scanning ion conductance microscopy (SICM), has been used to directly image membrane remodeling of CMs at different locations on the tissue surface. Periodic crest/valley patterns with a distance close to the sarcomere length appeared on the membrane of CMs near the edge of the tissue after ES, suggesting the enhanced transverse tubulation network. The SICM observation is also supported by the fluorescence images of the transverse tubulation network and α-actinin. Correspondingly, essential cardiac functions such as calcium handling and contraction force generation were improved. Our study provides evidence that chronic subthreshold ES can still improve the structural and functional developments of hECTs.

Calcium oscillation on homogeneous and heterogeneous networks of ryanodine receptor.

Calcium oscillation is an important calcium homeostasis, imbalance of which is the key mechanism of initiation and progression of many major diseases. The formation and maintenance of calcium homeostasis are closely related to the spatial distribution of calcium channels on endoplasmic reticulum, whose complex structure was unveiled by recent observations with superresolution imaging techniques. In the current paper, a theoretical framework is established by abstracting the spatial distribution of the calcium channels as a nonlinear biological complex network with calcium channels as nodes and Ca^{2+} as edges. A dynamical model for a ryanodine receptor (RyR) is adopted to investigate the effect of spatial distribution on calcium oscillation. The mean-field model can be well reproduced from the complete graph and dense Erdös-Rényi network. The synchronization of RyRs is found important to generate a global calcium oscillation. Below a critical density of the Erdös-Rényi or BaraBási-Albert network, the amplitude and interspike interval decrease rapidly with the end of disappearance of oscillation due to the desynchronization. The clique graph with a cluster structure cannot produce a global oscillation due to the failure of synchronization between clusters. A more realistic geometric network is constructed in a two-dimensional plane based on the experimental information about the RyR arrangement of clusters and the frequency distribution of cluster sizes. Different from the clique graph, the global oscillation can be generated with reasonable parameters on the geometric network. The simulation also suggests that existence of small clusters and rogue RyRs plays an important role in the maintenance of global calcium oscillation through keeping synchronization between large clusters. Such results support the heterogeneous distribution of RyRs with different-size clusters, which is helpful to understand recent observations with superresolution nanoscale imaging techniques. The current theoretical framework can also be extent to investigate other phenomena in calcium signal transduction.

Bulk and surface exsolution produces a variety of Fe-rich and Fe-depleted ellipsoidal nanostructures in La0.6Sr0.4FeO3 thin films.

The past several years have seen a resurgence in the popularity of metal exsolution as an approach to synthesize advanced materials proposed for novel catalytic, magnetic, optical, and electrochemical properties. Whereas most studies to-date have focused on surface exsolution (motivated by catalysis), we instead report on the diversity of nanostructures formed in La0.6Sr0.4FeO3 thin films during sub-surface or so-called 'bulk' exsolution, in addition to surface exsolution. Bulk exsolution is a promising approach to tuning the functionality of materials, yet there is little understanding of the nanostructures exsolved within the bulk and how they compare to those exsolved at gas-solid interfaces. This work combines atomic- and nano-scale imaging and spectroscopy techniques applied using a state-of-the-art aberration-corrected scanning transmission electron microscope (STEM). In doing so, we present a detailed atomic-resolution study of a range of Fe-rich and Fe-depleted nanostructures possible via exsolution, along with qualitative and quantitative chemical analysis of the exsolved nanostructures and oxide phases formed throughout the film. Local structural changes in the perovskite matrix, coinciding with nanostructure exsolution, are also characterized with atomic-resolution STEM imaging. Fe exsolution is shown to create local A-site rich domains of Ruddlesden-Popper phase, and some stages of this phase formation have been demonstrated in this work. In particular, phase boundaries are found to be the primary nucleation sites for bulk and surface exsolution, and the exsolved particles observed here tend to be ellipsoidal with shape factor of 1.4. We report a range of nanostructure types (core-shell, bulk core-shell, adjacent, and independent particles), revealing several possible avenues of future exploration aimed to understand the formation mechanism of each exsolution type and to develop their functionality. This work is thus relevant to materials scientists and engineers motivated to understand and utilize exsolution to synthesize materials with predictable nanostructures.

Dual energy X-ray beam ptycho-fluorescence imaging.

X-ray ptychography and X-ray fluorescence are complementary nanoscale imaging techniques, providing structural and elemental information, respectively. Both methods acquire data by scanning a localized beam across the sample. X-ray ptychography processes the transmission signal of a coherent illumination interacting with the sample, to produce images with a resolution finer than the illumination spot and step size. By enlarging both the spot and the step size, the technique can cover extended regions efficiently. X-ray fluorescence records the emitted spectra as the sample is scanned through the localized beam and its spatial resolution is limited by the spot and step size. The requisites for fast ptychography and high-resolution fluorescence appear incompatible. Here, a novel scheme that mitigates the difference in requirements is proposed. The method makes use of two probes of different sizes at the sample, generated by using two different energies for the probes and chromatic focusing optics. The different probe sizes allow to reduce the number of acquisition steps for the joint fluorescence-ptychography scan compared with a standard single beam scan, while imaging the same field of view. The new method is demonstrated experimentally using two undulator harmonics, a Fresnel zone plate and an energy discriminating photon counting detector.

Nanoscale imaging technique for accurate identification of brain tumor contour using NBDS method

Brain tumor identification is a difficult task in the processing of diagnostic images and a great deal of research is being performed. Normally, the doctor can evaluate their condition through an MRI scan for irregular brain tissue growth. In this research work, the Kaggle brain MRI database image is used. The boosted non-local means filter is used to reduce noise in the image acquired and to improve the quality of the proposed image technique. Identifying the exact location of the tumor is a challenge, and a novel segmentation method is proposed to evaluate the position of the tumor precisely here. Recently, the nanoscale imaging technique increases tumor detection sensitivity, accuracy, and classification into a benign situation. This segmentation activity is carried out using the nanotechnology detection system using our new region-based segmentation approach. The system suggested used for the identification of the brain tumor seed pixel in the range of nanoscale (nm) is the nanotechnology basis detection scheme. Ultimately, the 97.3% deep-neural network classification method helps to identify brain cancer by automatically extracting the function. The computation time was found to be less when compared to the existing method which is 12 s.

NanoSIMS Imaging of D/H Ratios on FIB Sections.

The D/H ratio imaging of weakly hydrated minerals prepared as focused ion beam (FIB) sections is developed in order to combine isotopic imaging by nanoscale secondary ion mass spectrometry (NanoSIMS) of micrometer-sized grains with other nanoscale imaging techniques, such as transmission electron microscopy. In order to maximize the accuracy, sensitivity, precision, and reproducibility of D/H ratios at the micrometer size, while minimizing the surface contamination at the same time, we explored all instrumental parameters known to influence the measurement of D/H ratios in situ. Optimal conditions were found to be obtained with the use of (i) a Cs+ ion source and detection of H- and D- at low mass resolving power, (ii) a primary beam intensity of 100 pA, and (iii) raster sizes in the range of 8-15 μm. Nominally anhydrous minerals were used to evaluate the detection limits and indicate a surface contamination level of ∼200 ppm equivalent H2O under these conditions. With the high primary intensity used here, the dwell time is not a parameter as critical as found in previous studies and a dwell time of 1 ms/px is used to minimize dynamic contamination during analysis. Analysis of FIB sections was found to reduce significantly static contamination due to sample preparation and improved accuracy compared to using polished sections embedded not only in epoxy but in indium as well. On amphiboles, the typical overall uncertainty including reproducibility is ∼20 ‰ on bulk FIB sections and ∼50 ‰ at the 1.5 μm scale using image processing (1σ).

Multimodal x-ray and electron microscopy of the Allende meteorite.

Multimodal microscopy that combines complementary nanoscale imaging techniques is critical for extracting comprehensive chemical, structural, and functional information, particularly for heterogeneous samples. X-ray microscopy can achieve high-resolution imaging of bulk materials with chemical, magnetic, electronic, and bond orientation contrast, while electron microscopy provides atomic-scale spatial resolution with quantitative elemental composition. Here, we combine x-ray ptychography and scanning transmission x-ray spectromicroscopy with three-dimensional energy-dispersive spectroscopy and electron tomography to perform structural and chemical mapping of an Allende meteorite particle with 15-nm spatial resolution. We use textural and quantitative elemental information to infer the mineral composition and discuss potential processes that occurred before or after accretion. We anticipate that correlative x-ray and electron microscopy overcome the limitations of individual imaging modalities and open up a route to future multiscale nondestructive microscopies of complex functional materials and biological systems.

Novel Technology for Lung Tumor Detection Using Nanoimage

Nowadays, nanotechnology is gaining more advantages and widely used in many real-life applications including minute tumor detection and effective diagnosis. Nanoscale imaging technique significantly increases the precision, accuracy of tumor detection, and classification of tumor into benign and malignant. Computerized tomography (CT) scan is important for lung cancer diagnosis and research, because it gives accurate segmentation results in lung tumor. In the proposed NBDS method, nanotechnology based detection scheme is used to detect the lung tumors in nanoscale range. Pulmonary nodules are symptoms of lung cancer. The shape and size of these pulmonary nodules are used to diagnose lung cancer in CT images. In the early stages, nodules are very small, and radiologist has to refer to many CT images to diagnose the disease, causing operator mistakes. Image processing algorithms are used as an aid to detect and localize nodules. Here, the input nanoimage is enhanced by using the technique of unsharp masking with the anisotropic filter. By using toboggan algorithm, lung cancer images are segmented. Nano measuring tool for graphical user interface is developed in MATLAB software to detect the lung tumor area or lung lesion in the body (in nanometers). Image classification and feature extraction are done by K-nearest neighbor (KNN) and support vector machine (SVM) with Bag of Visual Words (BoVW) classifiers. The overall accuracy of 97% is obtained using GLCM and FOS features in MATLAB software. The convolutional neural network (CNN) classifier gives the maximum accuracy of 98.19% in MATLAB 2017a software, and hence, this classifier does not need the feature extraction step. Furthermore, the average time consumption for one lesion segmentation was under 4 s using our proposed method. The self-assembled biocompatible nano technique is created to detect the tumor area (in nanometers), and it automatically evaluates the disease. This novel nanotechnology-based tumor area detection schemes achieve robust, efficient, and accurate lung lesion segmentation in nano CT images.

Visualizing Dark Microglia.

Dark microglia, a recently described phenotype, are found in high numbers in nonhomeostatic conditions (e.g., Alzheimer's disease pathology, aging, chronic stress). As a specific protein marker has not yet been defined, they cannot be studied using conventional cellular biology techniques. They are recognized by their unique ultrastructural features visible under electron microscopy. This nanoscale resolution imaging technique allows the identification of cells based on their ultrastructure or immunoreactivity to certain proteins. In this protocol, we describe the steps necessary for the preparation of high-quality brain tissues for transmission electron microscopy, the imaging, the identification of dark microglia, and the ultrastructural analysis of various parameters that can be studied in these cells.

Multipoint-projection x-ray microscopy

Point projection is a mature geometry of x-ray imaging that is implemented in scientific and industrial applications. Objects to be imaged are placed near a microscopic x-ray source, and the magnification is accomplished by x-ray propagation towards a distant detector. The source size is a trade-off between the signal level and the spatial resolution. In this work, we demonstrate multipoint-projection x-ray imaging realized with an x-ray tube and compound structured microcapillary optics that generates nearly one thousand submicrometer secondary x-ray sources. The generated microbeams are multiplexed at the object. Demultiplexing of the transmitted beams, magnification, and phase contrast are achieved by the free-space propagation. A massive improvement in the signal-to-noise ratio, relative to a single secondary source, was achieved without a loss in spatial resolution. Hence, x-ray projections of highly absorbing samples at submicrometer spatial resolution could be recorded with only a few photons per detector pixel. For weakly absorbing samples, the multipoint-projection method enabled us to record, in a parallel way, replicated in-line x-ray holograms with incoherent radiation from an x-ray tube. Our method may enrich the panel of x-ray nanoscale imaging techniques and may be adopted for on-chip x-ray photonic devices.

Cardiomyocyte Membrane Structure and cAMP Compartmentation Produce Anatomical Variation in β2AR-cAMP Responsiveness in Murine Hearts

SummaryCardiomyocytes from the apex but not the base of the heart increase their contractility in response to β2-adrenoceptor (β2AR) stimulation, which may underlie the development of Takotsubo cardiomyopathy. However, both cell types produce comparable cytosolic amounts of the second messenger cAMP. We investigated this discrepancy using nanoscale imaging techniques and found that, structurally, basal cardiomyocytes have more organized membranes (higher T-tubular and caveolar densities). Local membrane microdomain responses measured in isolated basal cardiomyocytes or in whole hearts revealed significantly smaller and more short-lived β2AR/cAMP signals. Inhibition of PDE4, caveolar disruption by removing cholesterol or genetic deletion of Cav3 eliminated differences in local cAMP production and equilibrated the contractile response to β2AR. We conclude that basal cells possess tighter control of cAMP because of a higher degree of signaling microdomain organization. This provides varying levels of nanostructural control for cAMP-mediated functional effects that orchestrate macroscopic, regional physiological differences within the heart.