Micrometer Resolution Research Articles

Laminography as a tool for imaging large-size samples with high resolution.

Despite the increased brilliance of the new generation synchrotron sources, there is still a challenge with high-resolution scanning of very thick and absorbing samples, such as a whole mouse brain stained with heavy elements, and, extending further, brains of primates. Samples are typically cut into smaller parts, to ensure a sufficient X-ray transmission, and scanned separately. Compared with the standard tomography setup where the sample would be cut into many pillars, the laminographic geometry operates with slab-shaped sections significantly reducing the number of sample parts to be prepared, the cutting damage and data stitching problems. In this work, a laminography pipeline for imaging large samples (>1 cm) at micrometre resolution is presented. The implementation includes a low-cost instrument setup installed at the 2-BM micro-CT beamline of the Advanced Photon Source. Additionally, sample mounting, scanning techniques, data stitching procedures, a fast reconstruction algorithm with low computational complexity, and accelerated reconstruction on multi-GPU systems for processing large-scale datasets are presented. The applicability of the whole laminography pipeline was demonstrated by imaging four sequential slabs throughout an entire mouse brain sample stained with osmium, in total generating approximately 12 TB of raw data for reconstruction.

A step towards 6D WAXD tensor tomography.

X-ray scattering/diffraction tensor tomography techniques are promising methods to acquire the 3D texture information of heterogeneous biological tissues at micrometre resolution. However, the methods suffer from a long overall acquisition time due to multi-dimensional scanning across real and reciprocal space. Here, a new approach is introduced to obtain 3D reciprocal information of each illuminated scanning volume using mathematic modeling, which is equivalent to a physical scanning procedure for collecting the full reciprocal information required for voxel reconstruction. The virtual reciprocal scanning scheme was validated by a simulated 6D wide-angle X-ray diffraction tomography experiment. The theoretical validation of the method represents an important technological advancement for 6D diffraction tensor tomography and a crucial step towards pervasive applications in the characterization of heterogeneous materials.

Stylize My Wrinkles: Bridging the Gap from Simulation to Reality

AbstractModeling realistic human skin with pores and wrinkles down to the milli‐ and micrometer resolution is a challenging task. Prior work showed that such micro geometry can be efficiently generated through simulation methods, or in specialized cases via 3D scanning of real skin. Simulation methods allow to highly customize the wrinkles on the face, but can lead to a synthetic look. Scanning methods can lead to a more organic look for the micro details, however these methods are only applicable to small skin patches due to the required image resolution. In this work we aim to overcome the gap between synthetic simulation and real skin scanning, by proposing a method that can be applied to large skin regions (e.g. an entire face) with the controllability of simulation and the organic look of real micro details. Our method is based on style transfer at its core, where we use scanned displacement maps of real skin patches as style images and displacement maps from an artist‐friendly simulation method as content images. We build a library of displacement maps as style images by employing a simplified scanning setup that can capture high‐resolution patches of real skin. To create the content component for the style transfer and to facilitate parameter‐tuning for the simulation, we design a library of preset parameter values depicting different skin types, and present a new method to fit the simulation parameters to scanned skin patches. This allows fully‐automatic parameter generation, interpolation and stylization across entire faces. We evaluate our method by generating realistic skin micro details for various subjects of different ages and genders, and demonstrate that our approach achieves a more organic and natural look than simulation alone.

Closed-Loop Optical Tracking of a Micro-Conveyor over a Smart Surface

In this work, a closed loop control system is developed to optically localize and track micro-robots with high precision. These micro-robots (i.e., micro-conveyors) are in motion simultaneously across a smart surface.The developed method’s primary objectives are to optimize their trajectories, avoid collisions between them, and control their position with micrometric resolution. This article presents and characterizes the tracking of a single micro-conveyor, and the method works similarly when multiple micro-robots move over the surface. Our tracking method starts with a scanning phase, where a 2D steering mirror, placed above the smart surface, reflects a laser beam toward the conveying surface seeking for the target. Localization occurs when this light beam reaches the micro-conveyor. By adding a retro-reflective element, that reflects the light in the same direction of the the incident light, onto the surface of the micro-conveyor, the light will be reflected towards a photodetector. Depending on the feedback from the photodetector, the steering mirror rotates to track the trajectory of the micro-conveyor. The tip-tilt angular values of the steering mirror allows the micro-conveyor position to be obtained via calibrated localization system. The aim of this work is to regulate the micro-conveyor, within a closed-loop control system, to reduce the positional error between the actual and desired position. The actual position value is measured in real-time application using our developed optical sensor. Results for tracking in the x-and y-axis have validated the proposed method, with an average tracking error less than 30 µm within a range 150 mm × 150 mm.

High-resolution metal 3D printing via digital light processing

Three-dimensional (3D) printing of complex metallic parts with a micrometer resolution can extend the application of additive manufacturing to various fields wherein the mechanical, thermal, and electrical properties of metals are essential. Digital light processing (DLP) offers high-resolution and scalable 3D printing via vat photopolymerization (VPP). We present a high-precision metal 3D printing method, namely lithography metal additive manufacturing (LMAM) that uses DLP for a photosensitive resin filled with metal powder. The printing process and sintering conditions for a stainless-steel feedstock are discussed. Highly complex and hollow structures with a spatial resolution of 35 μm and surface roughness as low as 1 <Ra < 2 μm can be printed on a decimeter scale without any support structures. The density and tensile strength of the sintered parts are 99.3 % and 93 %, respectively, of those of annealed 316 L steel. The LMAM method is suitable for manufacturing small and highly accurate devices with tailored designs in diverse areas, such as microheat exchangers, biomedical implants, and metamaterials, as well as extends metal 3D printing to chemical engineering for pharmaceutical applications.

In Situ Helium Isotope Microimaging of Meteorites.

Isotope imaging is commonly used to investigate the localization of trace elements and their isotopes. In situ noble gas analysis of meteorites revealed the distribution of primordial noble gases that were trapped in the building blocks of asteroids and planets during the early stage of the solar system evolution. Solar wind noble gases are among the primordial gases present in meteorites and were trapped through exposure to solar wind. Micrometer-resolution in situ noble gas analysis has not been achieved due to the lack of sensitivity and spatial resolution. The microscale imaging technique is crucial for identifying the carrier phase of the solar wind noble gases. We have developed 4He isotope imaging utilizing secondary neutral mass spectrometry with strong field postionization. This technique achieved a lateral resolution of 2 μm and a 4He detection limit of 2 × 1017 cm-3. This development allows for the study of a solar wind gas-rich meteorite, Northwest Africa 801 carbonaceous chondrite, with micrometer resolution. The solar wind 4He carriers are fine-grained particles and are sparsely scattered in the matrix region.

Deep learning-based vessel extraction in 3D confocal microscope images of cleared human glioma tissues.

Comprehensive visualization and accurate extraction of tumor vasculature are essential to study the nature of glioma. Nowadays, tissue clearing technology enables 3D visualization of human glioma vasculature at micron resolution, but current vessel extraction schemes cannot well cope with the extraction of complex tumor vessels with high disruption and irregularity under realistic conditions. Here, we developed a framework, FineVess, based on deep learning to automatically extract glioma vessels in confocal microscope images of cleared human tumor tissues. In the framework, a customized deep learning network, named 3D ResCBAM nnU-Net, was designed to segment the vessels, and a novel pipeline based on preprocessing and post-processing was developed to refine the segmentation results automatically. On the basis of its application to a practical dataset, we showed that the FineVess enabled extraction of variable and incomplete vessels with high accuracy in challenging 3D images, better than other traditional and state-of-the-art schemes. For the extracted vessels, we calculated vascular morphological features including fractal dimension and vascular wall integrity of different tumor grades, and verified the vascular heterogeneity through quantitative analysis.

3D models of the cardiac conduction system in healthy neonatal human hearts

Iatrogenic damage to the cardiac conduction system (CCS) remains a significant risk during congenital heart surgery. Current surgical best practice involves using superficial anatomical landmarks to locate and avoid damaging the CCS. Prior work indicates inherent variability in the anatomy of the CCS and supporting tissues. This study introduces high-resolution, 3D models of the CCS in normal pediatric human hearts to evaluate variability in the nodes and surrounding structures. Human pediatric hearts were obtained with an average donor age of 2.7 days. A pipeline was developed to excise, section, stain, and image atrioventricular (AVN) and sinus nodal (SN) tissue regions. A convolutional neural network was trained to enable precise multi-class segmentation of whole-slide images, which were subsequently used to generate high- resolution 3D tissue models. Nodal tissue region models were created. All models (10 AVN, 8 SN) contain tissue composition of neural tissue, vasculature, and nodal tissues at micrometer resolution. We describe novel nodal anatomical variations. We found that the depth of the His bundle in females was on average 304 μm shallower than those of male patients. These models provide surgeons with insight into the heterogeneity of the nodal regions and the intricate relationships between the CCS and surrounding structures.

Two‐Photon Laser Printing to Mechanically Stimulate Multicellular Systems in 3D

AbstractBiological activities take place in 3D environments, where cells interact in various directions in a defined, often microstructured, space. A sub‐millimeter‐sized stretching device is developed to mechanically stimulate a structurally restricted, soft multicellular microenvironment to investigate the effect of defined cyclic mechanical forces on a multicellular system. It consists of a multi‐material 3D microstructure made of Polydimethylsiloxane (PDMS) and gelatine‐based hydrogel, which is printed using the 2‐photon polymerization (2PP) method. The printed structures are first characterized microscopically and mechanically to study the effect of different printing parameters. Using 2PP, organotypic cell cultures are then directly printed into the hydrogel structures to create true 3D cell culture systems. These systems are mechanically stimulated with a cantilever by indenting at defined positions. The cells in the 3D organotypic cell culture change morphology and actin orientation when exposed to cyclic mechanical stretch, even within short timescales of 30 min. As proof of concept, a Medaka retinal organoid is encapsulated in the same structure to demonstrate that even preformed organoids can be stimulated by this method. The results highlight the capability of 2PP for manufacturing multifunctional soft devices to mechanically control multicellular systems at micrometer resolution and thus mimic mechanical stresses as they occur in vivo.

Femtosecond electron beam probe of ultrafast electronics

The need for ever-faster information processing requires exceptionally small devices that operate at frequencies approaching the terahertz and petahertz regimes. For the diagnostics of such devices, researchers need a spatiotemporal tool that surpasses the device under test in speed and spatial resolution. Consequently, such a tool cannot be provided by electronics itself. Here we show how ultrafast electron beam probe with terahertz-compressed electron pulses can directly sense local electro-magnetic fields in electronic devices with femtosecond, micrometre and millivolt resolution under normal operation conditions. We analyse the dynamical response of a coplanar waveguide circuit and reveal the impulse response, signal reflections, attenuation and waveguide dispersion directly in the time domain. The demonstrated measurement bandwidth reaches 10 THz and the sensitivity to electric potentials is tens of millivolts or −20 dBm. Femtosecond time resolution and the capability to directly integrate our technique into existing electron-beam inspection devices in semiconductor industry makes our femtosecond electron beam probe a promising tool for research and development of next-generation electronics at unprecedented speed and size.

Capillary Manganese Halide Needle-Like Array Scintillator with Isolated Light Crosstalk for Micro-X-Ray Imaging.

The exacerbation of inherent light scattering with increasing scintillator thickness poses a major challenge for balancing the thickness-dependent spatial resolution and scintillation brightness in X-ray imaging scintillators. Herein, a thick pixelated needle-like array scintillator capable of micrometer resolution is fabricated via waveguide structure engineering. Specifically, this involves integrating a straightforward low-temperature melting process of manganese halide with an aluminum-clad capillary template. In this waveguide structure, the oriented scintillation photons propagate along the well-aligned scintillator and are confined within individual pixels by the aluminum reflective cladding, as substantiated from the comprehensive analysis including laser diffraction experiments. Consequently, thanks to isolated light-crosstalk channels and robust light output due to increased thickness, ultrahigh spatial resolutions of 60.8 and 51.7 lp mm-1 at a modulation transfer function (MTF) of 0.2 are achieved on 0.5mm and even 1mm thick scintillators, respectively, which both exceed the pore diameter of the capillary arrays' template (Φ = 10µm). As far as it is known, these micrometer resolutions are among the highest reported metal halide scintillators and are never demonstrated on such thick scintillators. Here an avenue is presented to the demand for thick scintillators in high-resolution X-ray imaging across diverse scientific and practical fields.

Bioactive gelatin-sheets as novel biopapers to support prevascularization organized by laser-assisted bioprinting for bone tissue engineering

Despite significant advances in the management of patients with oral cancer, maxillofacial reconstruction after ablative surgery remains a clinical challenge. In bone tissue engineering, biofabrication strategies have been proposed as promising alternatives to solve issues associated with current therapies and to produce bone substitutes that mimic both the structure and function of native bone. Among them, laser-assisted bioprinting (LAB) has emerged as a relevant biofabrication method to print living cells and biomaterials with micrometric resolution onto a receiving substrate, also called ‘biopaper’. Recent studies have demonstrated the benefits of prevascularization using LAB to promote vascularization and bone regeneration, but mechanical and biological optimization of the biopaper are needed. The aim of this study was to apply gelatin-sheet fabrication process to the development of a novel biopaper able to support prevascularization organized by LAB for bone tissue engineering applications. Gelatin-based sheets incorporating bioactive glasses (BGs) were produced using various freezing methods and crosslinking (CL) parameters. The different formulations were characterized in terms of microstructural, physical, mechanical, and biological properties in monoculture and coculture. Based on multi-criteria analysis, a rank scoring method was used to identify the most relevant formulations. The selected biopaper underwent additional characterization regarding its ability to support mineralization and vasculogenesis, its bioactivity potential and in vivo degradability. The biopaper ‘Gel5wt% BG1wt%—slow freezing—CL160 °C 24 h′ was selected as the best candidate, due to its suitable properties including high porosity (91.69 ± 1.55%), swelling ratio (91.61 ± 0.60%), Young modulus (3.97 × 104 ± 0.97 × 104 Pa) but also its great cytocompatibility, osteogenesis and bioactivity properties. The preorganization of human umbilical vein endothelial cell using LAB onto this new biopaper led to the formation of microvascular networks. This biopaper was also shown to be compatible with 3D-molding and 3D-stacking strategies. This work allowed the development of a novel biopaper adapted to LAB with great potential for vascularized bone biofabrication.

Phase Aberration Correction for In Vivo Ultrasound Localization Microscopy Using a Spatiotemporal Complex-Valued Neural Network.

Ultrasound Localization Microscopy (ULM) can map microvessels at a resolution of a few micrometers ( [Formula: see text]). Transcranial ULM remains challenging in presence of aberrations caused by the skull, which lead to localization errors. Herein, we propose a deep learning approach based on recently introduced complex-valued convolutional neural networks (CV-CNNs) to retrieve the aberration function, which can then be used to form enhanced images using standard delay-and-sum beamforming. CV-CNNs were selected as they can apply time delays through multiplication with in-phase quadrature input data. Predicting the aberration function rather than corrected images also confers enhanced explainability to the network. In addition, 3D spatiotemporal convolutions were used for the network to leverage entire microbubble tracks. For training and validation, we used an anatomically and hemodynamically realistic mouse brain microvascular network model to simulate the flow of microbubbles in presence of aberration. The proposed CV-CNN performance was compared to the coherence-based method by using microbubble tracks. We then confirmed the capability of the proposed network to generalize to transcranial in vivo data in the mouse brain (n=3). Vascular reconstructions using a locally predicted aberration function included additional and sharper vessels. The CV-CNN was more robust than the coherence-based method and could perform aberration correction in a 6-month-old mouse. After correction, we measured a resolution of [Formula: see text] for younger mice, representing an improvement of 25.8%, while the resolution was improved by 13.9% for the 6-month-old mouse. This work leads to different applications for complex-valued convolutions in biomedical imaging and strategies to perform transcranial ULM.

Scaling laws for near-wall flows of thixo-elasto-viscoplastic fluids in a millifluidic channel

Thixo-elasto-viscoplastic (TEVP) fluids are very complex fluids. In addition to elasticity and viscoplasticity, they exhibit thixotropy, i.e., time-dependent rheology due to breakdown and recovery of internal structures at different length- and timescales. General and consistent methods for a priori flow prediction of TEVP fluids based on rheological characteristics are yet to be developed. We report a combined study of the rheology and flow of 18 samples of different TEVP fluids (three yogurts and three concentrations of Laponite and Carbopol, respectively, in water in both the unstirred and a stirred state). The rheology is determined both with standard protocols and with an ex situ protocol aiming at reproducing the shear history of the fluid in the flow. Micrometer resolution flow measurements in a millimeter scale rectangular duct are performed with Doppler Optical Coherence Tomography (D-OCT). As expected, the results show the existence of a plug flow region for samples with sufficiently high yield stress. At low flow rates, the plug extends almost all the way to the wall and the extent of the plug decreases not only with increased flow rate but also with increased thixotropy. The ex situ rheology protocol enables estimation of the shear rate and shear stress close to the wall, making it possible to identify two scaling laws that relates four different non-dimensional groups quantifying the key properties wall-shear stress and slip velocity. The scaling laws are suggested as an ansatz for a priori prediction of the near-wall flow of TEVP fluids based on shear flow-curves obtained with a rheometer.

A low-cost close-range photogrammetric surface scanner

IntroductionA low-cost, close-range photogrammetric surface scanner is proposed, made from Computer Numerical Control (CNC) components and an off-the-shelf, consumer-grade macro camera.MethodsTo achieve micrometer resolution in reconstruction, accurate and photorealistic surface digitization, and retain low manufacturing cost, an image acquisition approach and a reconstruction method are proposed. The image acquisition approach uses the CNC to systematically move the camera and acquire images in a grid tessellation and at multiple distances from the target surface. A relatively large number of images is required to cover the scanned surface. The reconstruction method tracks keypoint features to robustify correspondence matching and uses far-range images to anchor the accumulation of errors across a large number of images utilized.Results and discussionQualitative and quantitative evaluation demonstrate the efficacy and accuracy of this approach.

First experimental evaluation of count-rate performance for micrometre resolution deep silicon detector

Objective. An ultra-fine-pitch deep silicon detector has been developed for clinical photon-counting computed tomography (CT). With a small pixel size of 14 × 650 μm2, it has shown potential to reach micrometre spatial resolution in previous simulation studies. A detector prototype with such geometry has been manufactured, and we report on the first experimental evaluation of its count-rate performance. Approach. The measurement was carried out at MAX IV synchrotron laboratory with 35 keV monochromatic x-rays. By inserting tungsten attenuators of 50, 75, 100, 150, 200, 225, 325 μ m-thicknesses into the beam, the response of the detector to fluence rates from 3.3 × 107 to 1.3 × 1011 mm−2 s−1 was characterized. Main results. The measurement result showed that the detector exhibited count rate linearity up to 6.66 × 108 mm−2 s−1 with 13% count loss and was still functional at count rate up to 2.9 × 1010 mm−2 s−1. A semi-nonparalyzable dead-time model was fitted to the count-rate behaviour of the detector, showing great agreement with the measured data, with an estimated nonparalyzable dead time of 2.9 ns. Significance. This is the first experimental evaluation of the count-rate performance for a deep silicon detector with such small pixel geometry. The results suggest that this type of detector shows the potential to be used at fluence rates encountered in clinical CT with little count loss due to pile-up.

Non-Invasive High-Resolution Imaging of In Vivo Human Myelinated Axons

This work aims to reveal the microscopic (2–3 micrometer resolution) appearance of human myelinated nerve fibers in vivo for the first time. We analyzed the myelinated retinal nerve fibers of a male patient without other neurological disorders in a non-invasive way using the transscleral optical phase imaging method with adaptive optics. We also analyzed the fellow eye with non-myelinated nerve fibers and compared the results with traditional ocular imaging methods such as optical coherence tomography. We documented the microscopic appearance of human myelin and myelinated axons in vivo. This method allowed us to obtain better details than through traditional ocular imaging methods. We hope these findings will be useful to the scientific community to evaluate neuro-retinal structures through new imaging techniques and more accurately document nerve anatomy and the pathophysiology of this disease.

Transparent and Planar Cs3Cu2Cl5 Crystals for Micrometer-Resolution X-ray Imaging Screen.

Copper-based (I) halide perovskites have emerged as a promising candidate for scintillation screens in X-ray inspection and imaging areas due to their solution processability and high light yield. Here, a centimeter-sized Cs3Cu2Cl5 single crystal was grown by a slow-cooling method. The planar orientation was controlled in a space-confined chamber, generating a planar crystal which is readily used for a scintillation screen without any further shaping. The crystal exhibited a unity photoluminescence quantum yield and superior scintillation performance. The Cs3Cu2Cl5 single crystal exhibited a high light yield up to 95,000 photons/MeV, which enabled an X-ray detector of a detection limit down to 2.7 μGyair/s. The homemade imager demonstrated a spatial resolution of 105 lp/mm, representing an unprecedented micrometer resolution in laboratory. Importantly, the stability of Cs3Cu2Cl5 was significantly improved by a new surface passivation procedure, whereby the passivated crystal reserved its phase after 6 months' storage in a vial. This work introduced a new solution-based synthetic method for two-dimensional scintillating crystals, opening many avenues to high-performance X-ray imaging applications.

Focused‐beam X‐ray fluorescence and diffraction microtomographies for mineralogical and chemical characterization of unsectioned extraterrestrial samples

AbstractThis study describes the application of new synchrotron X‐ray fluorescence (XRF) and diffraction (XRD) microtomographies for the 3‐D visualization of chemical and mineralogical variations in unsectioned extraterrestrial samples. These improved methods have been applied to three compositionally diverse chondritic meteorite samples that were between 300 and 400 μm in diameter, including samples prepared from fragments of the CR2 chondrite LaPaz Icefield (LAP) 02342, H5 chondrite MacAlpine Hills (MAC) 88203, and the CM2 chondrite Murchison. The synchrotron‐based XRF and XRD tomographies used are focused‐beam techniques that measure the intensities of fluorescent and diffracted X‐rays in a sample simultaneously during irradiation by a high‐energy microfocused incident X‐ray beam. Measured sinograms of the emitted and diffracted intensities were then tomographically reconstructed to generate 2‐D slices of XRF and XRD intensity through the sample, with reconstructed pixel resolution of 1–2 μm, defined by the resolution of the focused incident X‐ray beam. For sample LAP 02342, primary mineral phases that were visualized in reconstructed slices using these techniques included isolated grains of α‐Fe, orthopyroxene, and olivine. For our sample of MAC 88203, XRF/XRD tomography allowed visualization of forsteritic olivine as a primary mineral phase, a vitrified fusion crust at the sample surface, identification of localized Cr‐rich spinels at spatial resolutions of several micrometers, and imaging of a plagioclase‐rich glassy matrix. In the sample of Murchison, major identifiable phases include clinoenstatite‐ and olivine‐rich chondrules, variable serpentine matrix minerals and small Cr‐rich spinels. Most notable in the tomographic analysis of Murchison is the ability to quantitatively distinguish and visualize the complex mixture of serpentine‐group minerals and associated tochilinite–cronstedtite intergrowths. These methods provide new opportunities for spatially resolved characterization of sample texture, mineralogy, crystal structure, and chemical state in unsectioned samples. This provides researchers an ability to characterize such samples internally with minimal disruption of sample micro‐structures and chemistry, possibly without the need for sample extraction from some types of sampling and capture media.

Magnetic Resonance Imaging-Based Assessment of Pancreatic Fat Strongly Correlates With Histology-Based Assessment of Pancreas Composition.

The aim of the study is to assess the relationship between magnetic resonance imaging (MRI)-based estimation of pancreatic fat and histology-based measurement of pancreatic composition. In this retrospective study, MRI was used to noninvasively estimate pancreatic fat content in preoperative images from high-risk individuals and disease controls having normal pancreata. A deep learning algorithm was used to label 11 tissue components at micron resolution in subsequent pancreatectomy histology. A linear model was used to determine correlation between histologic tissue composition and MRI fat estimation. Twenty-seven patients (mean age 64.0 ± 12.0 years [standard deviation], 15 women) were evaluated. The fat content measured by MRI ranged from 0% to 36.9%. Intrapancreatic histologic tissue fat content ranged from 0.8% to 38.3%. MRI pancreatic fat estimation positively correlated with microanatomical composition of fat (r = 0.90, 0.83 to 0.95], P < 0.001); as well as with pancreatic cancer precursor ( r = 0.65, P < 0.001); and collagen ( r = 0.46, P < 0.001) content, and negatively correlated with pancreatic acinar ( r = -0.85, P < 0.001) content. Pancreatic fat content, measurable by MRI, correlates to acinar content, stromal content (fibrosis), and presence of neoplastic precursors of cancer.