Micrometer Resolution Research Articles

Modular Plasmonic Nanopore for Opto‐Thermal Gating

AbstractSolid‐state nanopore gating inspired by biological ion channels is gaining increasing traction due to a large range of applications in biosensing and drug delivery. Integration of stimuli‐responsive molecules such as poly(N‐isopropylacrylamide) (PNIPAM) inside nanopores can enable temperature‐dependent gating, which so far has only been demonstrated using external heaters. In this work, plasmonic resonators are combined inside the nanopore architecture with PNIPAM to enable optical gating of individual or multiple nanopores with micrometer resolution and a switching speed of a few milliseconds by thermo‐plasmonics effects. A temperature change of 40 kelvin per millisecond is achieved and demonstrates the efficacy of this method using nanopore ionic conductivity measurements that enable selective activation of individual nanopores in an array. Moreover, the selective gating of specific nanopores in an array can set distinct ionic conductance levels: low, medium, and high (i.e., “0,” “1,” and “2”), which can be exploited for logical gating with optical signal control. Such selective optical gating in nanopore arrays marks a breakthrough in nanofluidics, as it paves the way toward smart devices that offer multifunctional applications including biosensing, targeted drug delivery, and fluidic mixing.

Synchrotron X-ray imaging of soft biological tissues - principles, applications and future prospects.

Synchrotron-based tomographic phase-contrast X-ray imaging (SRµCT or SRnCT) is a versatile isotropic three-dimensional imaging technique that can be used to study biological samples spanning from single cells to human-sized specimens. SRµCT and SRnCT take advantage of the highly brilliant and coherent X-rays produced by a synchrotron light source. This enables fast data acquisition and enhanced image contrast for soft biological samples owing to the exploitation of phase contrast. In this Review, we provide an overview of the basics behind the technique, discuss its applications for biologists and provide an outlook on the future of this emerging technique for biology. We introduce the latest advances in the field, such as whole human organs imaged with micron resolution, using X-rays as a tool for virtual histology and resolving neuronal connections in the brain.

Magnetic resonance microscopy for submillimeter samples in a horizontal MR scanner

The spatial resolution in magnetic resonance imaging is mainly limited by low SNR, which is commonly addressed by long measurement times or dedicated hardware. In single digit micron resolutions, diffusion becomes a further limiting factor since depending on the gradient strength, the diffusion length of particles may approach the target resolutions. Spatial resolution improvement has been addressed by microscopy inserts comprising dedicated gradient systems and RF-coils, usually designed for NMR spectrometers that are often equipped with a deuterium lock for field drift compensations. The presented microscopy insert has been designed to provide single-digit micron resolutions on horizontal preclinical imaging systems utilizing their full imaging and user interface capabilities. The incorporated gradient provides an efficiency of 0.135 T/(m∙A) which in combination with the system’s gradient amplifiers yields a maximum of 27 T/m. With the additional low noise amplifier added to the RF-path a three-fold SNR improvement could be achieved for small samples. Furthermore, a modified constant time imaging sequence was introduced to improve the capability of the setup for ultra‐high-resolution imaging demonstrated on zebrafish embryos at different development stages with (9 μm)³ resolution.

Optical Mapping of Local Threshold Voltage With Micrometer Resolution in AlGaN/GaN HEMTs

Optical Mapping of Local Threshold Voltage With Micrometer Resolution in AlGaN/GaN HEMTs

Thermal characterization of vertical interface by scanning photothermal radiometry.

In this work, scanning photothermal radiometry is used for imaging and to characterize a submicron crack. From the thermal images, the evolution of the crack is mapped in the space with micrometer resolution. A vertical contact interface at the steel-steel junction is used to represent a micro-crack with a thickness less than 0.5μm. The thermal quadrupole approach is used to model the heat transfer within the semi-infinite vertical crack. Then, using the phase mapping and that calculated from the model, the estimation of both the equivalent thermal boundary resistance of the interface and the average interface thickness was done.

Study of chemiluminescence of methane–air flame stabilized on a flat porous burner

In this work, the spatial distribution and spectral characteristics of the chemiluminescence of chemically excited species, OH∗ and CH∗, are experimentally and numerically studied by using a stationary premixed methane–air flame stabilized on the surface of a flat porous burner for various equivalence ratio and normal pressure. Numerical simulations are carried out using detailed reaction mechanisms, and the experimental study includes high-resolution spatial and spectral optical measurements. Despite the data reported in the literature, it is found that (i) the rotational degrees of freedom of OH∗ and CH∗ are not in thermal equilibrium with the surrounding gas and therefore cannot be used to measure flame temperature; (ii) there is no direct correlation between the heat release rate and the distribution of OH∗ and CH∗; (iii) the detailed reaction mechanisms not only quantitatively, and also qualitatively differ in description of the OH∗ and CH∗ concentrations. Since the chemically excited species are well localized in a direction normal to the flame surface, they are demonstrated to be a very accurate markers of flame location. The shape of the combustion front can be reconstructed and resolved up to the accuracy of tens of microns, which is very important for estimation of blow-off critical parameters and measurement of the laminar burning velocity.Novelty and significance statementCurrently, there is a growing interest in the development of sensors for combustion control systems, including active control and suppression of instabilities, in combustion chambers of various devices and engines based on chemiluminescence of excited reaction species. The possibility of non-invasive determination of parameters such as flame temperature, stoichiometry, heat release rate location, etc. using this technique is discussed. We have found that most of these parameters cannot be estimated either due to fundamental limitations or insufficient knowledge of the reaction kinetics involved in the production of these species. Nevertheless, since OH* and CH* are well localized in the direction normal to the flame surface, they can be used as very accurate markers of flame shape and position, allowing us to reconstruct the flame surface to within tens of microns resolution, which is very important for estimating blow-off critical parameters and measuring laminar burning velocity.

Syneresis-Driven Self-Refilling Printing of Geometry/Component-Controlled Nano/Microstructures.

Nano/microfabrication is of fundamental importance both in scientific and industrial situations. There are, therefore, many attempts at realizing easier, quicker, and more precise fabrication of various structures; however, achieving this aim without a bulky and costly setup is still challenging. Here, we introduce a facile and versatile means of printing an ordered structure consisting of nanoscale stripes and more complicated geometries including pillars and wavy form with a lateral resolution of single micrometers. To this end, we prepare a polydimethylsiloxane (PDMS) slab with an oxygen plasma-induced wrinkled surface where liquid PDMS exudes by syneresis. Since this liquid PDMS is automatically loaded, the printing is repeatable without inking. A substrate moderately wettable to the liquid PDMS as well as amount/property-controlled syneresis is primarily important for the creation of well-defined structures. Precisely controlling these conditions will make this method universally applicable to diverse substrates and liquids including suspensions.

High-Precision Printing of Cermets by Collapsable Matrix Assisted Digital Light Processing.

Shaping hard and brittle materials, e.g. cermets, at micrometer resolution has long been known challenging for both mechanical machining and high energy beam based additive manufacturing. Digital light processing (DLP), which features great printing quality and decent precision, unfortunately lacks capability to deal with the popular slurry-typed cermet precursor due to the tremendous optical absorption by its particles. Here, an innovative protocol based on a versatile collapsable matrix is devised to allow high-precision printing of WC-Co cermets on DLP platform. By tuning the external environment, this matrix attenuates composite powders to facilitate photopolymerization at the printing stage, and shrinks to condense green parts prior to thermal sintering. The as-obtained samples by collapsable matrix assisted DLP can reach a relative density of ≈90%, a record-breaking resolution of ≈10µm, and a microhardness of up to 14.5GPa. Complex delicate structures, including school emblem, honeycomb, and micro-drill can be directly fabricated, which has never been achieved before. Impressively, the as-obtained micro-drill is able to be directly used in drilling tasks. The above strategy represents a great progress in DLP by enabling shaping strong light attenuating materials at high resolution. Such advantages are ideal for the next generation ceramic-metal composite additive manufacturing.

Robotic Ultrasound and Novel Collagen Analyses for Polycystic Kidney Disease Research Using Mice.

3D imaging and histology are critical tools for assessing polycystic kidney disease ( PKD ) in patients and animal models. Magnetic resonance ( MR ) imaging provides micron resolution, but is time consuming, expensive, and access to equipment and expertise is limiting. Robotic ultrasound ( US ) imaging has lower spatial resolution but is faster, more cost effective, and accessible. Similarly, Picrosirius red ( PSR ) staining and brightfield microscopy is commonly used to assess fibrosis; however, alternative methods have been shown in non-kidney tissues to provide greater sensitivity and more detailed structural characterization. In this study, we evaluated the utility of robotic US and alternative methods of quantifying PSR staining for PKD research. We compared longitudinal total kidney volume ( TKV ) measurements using US and MR. We additionally compared PSR imaging and quantification using standard brightfield with that by circularly polarized light with hue analysis, and fluorescence imaging analyzed using CT-FIRE software for automatic detection of individual collagen fibers. Increased TKV was detected by US in Pkd1RC/RC vs wild type ( WT ) at timepoints spanning early to established disease. US inter-observer variability was greater but allowed scanning in 2-5 minutes/mouse while MR required 20-30 minutes/mouse. While no change in fibrotic index was detected in this cohort of relatively mild disease using brightfield, polarized light showed fibers skewed thinner in Pkd1RC/RC vs WT. Fluorescence imaging showed a higher density of collagen fibers in Pkd1RC/RC vs WT, and fibers were thinner and curvier with no change in length. Additionally, fiber density was higher in both glomeruli and tubules in Pkd1RC/RC , and glomeruli had a higher fiber density than tubules in Pkd1RC/RC , and trended higher in WT. These studies show robotic ultrasound is a rigorous imaging tool for pre-clinical PKD research. Additionally, they demonstrate the increased sensitivity of polarized and fluorescence analysis of PSR-stained collagen.

Three-dimensional fiber orientation mapping of the human brain at micrometer resolution.

The accurate measurement of three-dimensional (3D) fiber orientation in the brain is crucial for reconstructing fiber pathways and studying their involvement in neurological diseases. Comprehensive reconstruction of axonal tracts and small fascicles requires high-resolution technology beyond the ability of current in vivo imaging (e.g. diffusion magnetic resonance imaging). Optical imaging methods such as polarization-sensitive optical coherence tomography (PS-OCT) and polarization microscopy can quantify fiber orientation at micrometer resolution but have been limited to two-dimensional in-plane orientation or thin slices, preventing the comprehensive study of connectivity in 3D. In this work we present a novel method to quantify volumetric 3D orientation in full angular space with PS-OCT. We measure the polarization contrasts of the brain sample from two illumination angles of 0 and 15 degrees and apply a computational method that yields the 3D optic axis orientation and true birefringence. We further present 3D fiber orientation maps of entire coronal cerebrum sections and brainstem with 10 μm in-plane resolution, revealing unprecedented details of fiber configurations. We envision that our method will open a promising avenue towards large-scale 3D fiber axis mapping in the human brain as well as other complex fibrous tissues at microscopic level.

Ex vivo optical coherence tomography combined with near infrared targeted fluorescence: towards in-vivo esophageal cancer detection.

Early detection of (pre)malignant esophageal lesions is critical to improve esophageal cancer morbidity and mortality rates. In patients with advanced esophageal adenocarcinoma (EAC) who undergo neoadjuvant chemoradiation therapy, the efficacy of therapy could be optimized and unnecessary surgery prevented by the reliable assessment of residual tumors after therapy. Optical coherence tomography (OCT) provides structural images at a (sub)-cellular level and has the potential to visualize morphological changes in tissue. However, OCT lacks molecular imaging contrast, a feature that enables the study of biological processes at a cellular level and can enhance esophageal cancer diagnostic accuracy. We combined OCT with near-infrared fluorescence molecular imaging using fluorescently labelled antibodies (immuno-OCT). The main goal of this proof of principle study is to investigate the feasibility of immuno-OCT for esophageal cancer imaging. We aim to assess whether the sensitivity of our immuno-OCT device is sufficient to detect the tracer uptake using an imaging dose (∼100 times smaller than a dose with therapeutic effects) of a targeted fluorescent agent. The feasibility of immuno-OCT was demonstrated ex-vivo on dysplastic lesions resected from Barrett's patients and on esophageal specimens resected from patients with advanced EAC, who were respectively topically and intravenously administrated with the tracer bevacizumab-800CW. The detection sensitivity of our system (0.3 nM) is sufficient to detect increased tracer uptake with micrometer resolution using an imaging dose of labelled antibodies. Moreover, the absence of layered structures that are typical of normal esophageal tissue observed in OCT images of dysplastic/malignant esophageal lesions may further aid their detection. Based on our preliminary results, immuno-OCT could improve the detection of dysplastic esophageal lesions.

Individually Addressed Quantum Gate Interactions Using Dynamical Decoupling

A leading approach to implementing small-scale quantum computers has been to use laser beams, focused to micron spot sizes, to address and entangle trapped ions in a linear crystal. Here we propose a method to implement individually addressed entangling gate interactions, but driven by microwave fields, with a spatial resolution of a few microns, corresponding to 10−5 microwave wavelengths. We experimentally demonstrate the ability to suppress the effect of the state-dependent force using a single ion, and find the required interaction introduces 3.7(4)×10−4 error per emulated gate in a single-qubit benchmarking sequence. We model the scheme for a 17-qubit ion crystal, and find that any pair of ions should be addressable with an average crosstalk error of approximately 10−5. Published by the American Physical Society 2024

Image post-processing for SILMAS: structured illumination light sheet microscopy with axial sweeping.

In this article, we propose a post-processing scheme for the novel volumetric microscopy technique SILMAS. We demonstrate this scheme on data from an alpha-synuclein transgenic mouse brain. By combining structured illumination and axial sweeping, a SILMAS measurement provides a prerequisite for quantitative data extraction through improved contrast and optical sectioning. However, due to the technique's efficient removal ofb multiple scattered light, image artifacts such as illumination inhomogeneity, shadowing stripes, and signal attenuation, are highlighted in the recorded volumes. To suppress these artifacts, we rely on the strengths of the imaging method. The SILMAS data, together with the Beer-Lambert law, allow for an approximation of real light extinction, which can be used to compensate for light attenuation in a near-quantitative way. Shadowing stripes can be suppressed efficiently using a computational strategy thanks to the large numerical aperture of an axially swept light sheet. Here, we build upon prior research that employed wavelet-Fourier filtering by incorporating an extra bandpass step. This allows us to filter high-contrast light sheet microscopy data without introducing new artifacts and with minimal distortion of the data. The combined technique is suitable for imaging cleared tissue samples of up to a centimeter scale with an isotropic resolution of a few microns. The combination of a thin and uniform light sheet, scattered light suppression, light attenuation compensation, and shadowing suppression produces volumetric data that is seamless and highly uniform.

Imaging of developing human brains with ex vivo PSOCT and dMRI.

The human brain undergoes substantial developmental changes in the first five years of life. Particularly in the white matter, myelination of axons occurs near birth and continues at a rapid pace during the first 2 to 3 years. Diffusion MRI (dMRI) has revolutionized our understanding of developmental trajectories in white matter. However, the mm-resolution of in vivo techniques bears significant limitation in revealing the microstructure of the developing brain. Polarization sensitive optical coherence tomography (PSOCT) is a three-dimensional (3D) optical imaging technique that uses polarized light interferometry to target myelinated fiber tracts with micrometer resolution. Previous studies have shown that PSOCT contributes significantly to the elucidation of myelin content and quantification of fiber orientation in adult human brains. In this study, we utilized the PSOCT technique to study developing brains during the first 5 years of life in combination with ex vivo dMRI. The results showed that the optical properties of PSOCT quantitatively reveal the myelination process in young children. The imaging contrast of the optic axis orientation is a sensitive measure of fiber orientations in largely unmyelinated brains as young as 3-months-old. The micrometer resolution of PSOCT provides substantially enriched information about complex fiber networks and complements submillimeter dMRI. This new optical tool offers great potential to reveal the white matter structures in normal neurodevelopment and developmental disorders in unprecedented detail.

Off-axis reflective microscope objective with a centimeter scale field of view and micron resolution

Microscope objectives with wide field-of-view (FOV) and high resolution are urgently needed for the frontier research in life sciences. However, traditional transmission microscope objectives typically have a narrow FOV and severe chromatic aberration. A new off-axis reflective microscope objective with a centimeter scale FOV and micron resolution is proposed in this paper. This objective, with its simple structure, can operate over a wide wavelength range. A design method for a wide FOV optical system is presented, which can eliminate the obstruction of the rays and control the intermediate image plane. Using this method, we design a novel off-axis four-mirror microscope objective with a FOV of 10 mm × 1.5 mm and a numerical aperture of 0.33.

Scanning‐Laser‐Based Microstereolithography of Microfluidic Chips with Micron Resolution

AbstractThe constant improvement of stereolithography (SL) in terms of achievable resolution and printing time has sparked high expectations that SL will enable the rapid prototyping of truly microfluidic chips with features below 100 µm. However, most commercial high‐resolution stereolithography devices are based on Digital Light Processing (DLP) and thus sacrifice lateral printing size for resolution. Consequently, even 10 years after the advent of microstereolithography there is no commercialized 3D printing system that can effectively fulfill all the demands to replace soft lithography for microfluidic prototyping. In this work, for the first time, This study demonstrates that a commercial laser‐based stereolithography device is capable of manufacturing microfluidic chips with embedded channels smaller than 100 µm with a footprint of 7.24 × 0.3 cm2. A chip fabricated in poly(ethylene glycol) diacrylate (PEGDA) that can readily be used for fluid mixing, is presented in this study. This research shows that the accessibility of high‐resolution chips with footprints of several cm2, using laser‐based stereolithography, enables the manufacturing of truly microfluidic systems with high impact on prototyping and manufacturing.

3D imaging and flow analysis of sandstone pore structure

Abstract Micro CT technology can provide three-dimensional high-precision digital images with micrometer (or smaller) resolution, offering robust technical support for the study of the internal structure of rocks. Based on the data obtained from the CT scanning of Leopard sandstone, a series of image processes were carried out using the Avizo software to extract pore distribution. The watershed algorithm was employed to independently segment each pore, and the largest sphere algorithm was used to establish a sphere-tube model. Based on the constructed pore network model, quantitative analyses of the porosity, fractal dimensions, permeability, and other parameters of the sandstone sample were conducted. Moreover, the connectivity of the pores was analyzed using an automatic skeleton model. Results show: ① CT scanning digital core technology can not only visualize the three-dimensional connectivity and isolated pores of sandstone but also quantitatively characterize the pore structure parameters; ② The analyzed Leopard sandstone has a connected porosity of 13.76%, an isolated porosity of 0.53%, and the pore radius is mainly concentrated in the 0-70μm small pore range, occupying 76.1% of the total pore volume; ③ The lower the sample porosity, the higher the fractal dimension, indicating a more complex pore structure.

Gold-Hydrogel Nanocomposites for High-Resolution Laser-Based 3D Printing of Scaffolds with SERS-Sensing Properties.

Although visible light-based stereolithography (SLA) represents an affordable technology for the rapid prototyping of 3D scaffolds for in vitro support of cells, its potential could be limited by the lack of functional photocurable biomaterials that can be SLA-structured at micrometric resolution. Even if innovative photocomposites showing biomimetic, bioactive, or biosensing properties have been engineered by loading inorganic particles into photopolymer matrices, main examples rely on UV-assisted extrusion-based low-resolution processes. Here, SLA-printable composites were obtained by mixing a polyethylene glycol diacrylate (PEGDA) hydrogel with multibranched gold nanoparticles (NPs). NPs were engineered to copolymerize with the PEGDA matrix by implementing a functionalization protocol involving covalent grafting of allylamine molecules that have C═C pendant moieties. The formulations of gold nanocomposites were tailored to achieve high-resolution fast prototyping of composite scaffolds via visible light-based SLA. Furthermore, it was demonstrated that, after mixing with a polymer and after laser structuring, gold NPs still retained their unique plasmonic properties and could be exploited for optical detection of analytes through surface-enhanced Raman spectroscopy (SERS). As a proof of concept, SERS-sensing performances of 3D printed plasmonic scaffolds were successfully demonstrated with a Raman probe molecule (e.g., 4-mercaptobenzoic acid) from the perspective of future extensions to real-time sensing of cell-specific markers released within cultures. Finally, biocompatibility tests preliminarily demonstrated that embedded NPs also played a key role by inducing physiological cell-cytoskeleton rearrangements, further confirming the potentialities of such hybrid nanocomposites as groundbreaking materials in laser-based bioprinting.

X-ray scattering based scanning tomography for imaging and structural characterization of cellulose in plants.

X-ray and neutron scattering have long been used for structural characterization of cellulose in plants. Due to averaging over the illuminated sample volume, these measurements traditionally overlooked the compositional and morphological heterogeneity within the sample. Here, a scanning tomographic imaging method is described, using contrast derived from the X-ray scattering intensity, for virtually sectioning the sample to reveal its internal structure at a resolution of a few micrometres. This method provides a means for retrieving the local scattering signal that corresponds to any voxel within the virtual section, enabling characterization of the local structure using traditional data-analysis methods. This is accomplished through tomographic reconstruction of the spatial distribution of a handful of mathematical components identified by non-negative matrix factorization from the large dataset of X-ray scattering intensity. Joint analysis of multiple datasets, to find similarity between voxels by clustering of the decomposed data, could help elucidate systematic differences between samples, such as those expected from genetic modifications, chemical treatments or fungal decay. The spatial distribution of the microfibril angle can also be analyzed, based on the tomographically reconstructed scattering intensity as a function of the azimuthal angle.

A multi-electrode two-dimensional position sensitive diamond detector

In multi-electrode devices, charge pulses at all the electrodes are induced concurrently by the motion of the excess charge carriers generated by a single ion. This charge-sharing effect is such that the pulse amplitude at each sensitive electrode depends on the device geometry, its overall electrostatic configuration, and the charge transport properties of the detecting material. Therefore, the cross-analysis of the charge pulses induced at each electrode offers implicit information on the position of the ion impact. In this work, we investigate the two-dimensional position sensitivity of a diamond detector fabricated by deep ion beam lithography. By exploiting the ion beam induced charge technique, the device was exposed to a 2 MeV Li+ ion micro-beam to map the spatial dependence of the charge collection efficiency (CCE) on the nominal micro-beam scanning position. The combination of the CCE maps revealed a two-dimensional position sensitivity of the device with micrometric resolution at the center of the active region.