Investigation of an Active Focusing Planar Piezoelectric Ultrasonic Transducer.

Ultrasonic focusing transducers have broad prospects in advanced ultrasonic non-destructive testing fields. However, conventional focusing methods that use acoustic concave lenses can disrupt the acoustic impedance matching condition, thereby adversely affecting the sensitivity of the transducers. In this paper, an active focusing planar ultrasonic transducer is designed and presented to achieve a focusing effect with a higher sensitivity. An electrode pattern consisting of multiple concentric rings is designed, which is inspired by the structure of Fresnel Zone Plates (FZP). The structural parameters are optimized using finite element simulation methods. A prototype of the transducer is manufactured with electrode patterns made of conductive silver paste using silk screen-printing technology. Conventional focusing transducers using an acoustic lens and an FZP baffle are also manufactured, and their focusing performances are comparatively tested. The experimental results show that our novel transducer has a focal length of 16 mm and a center frequency of 1.16 MHz, and that the sensitivity is improved by 23.3% compared with the conventional focusing transducers. This research provides a new approach for the design of focusing transducers.

Possibility of using the imaged capillary isoelectric method as a multi-attribute method for bispecific antibodies.

An imaged capillary isoelectric focusing (icIEF)-based method was developed and validated as a multi-attribute method for a bispecific antibody (BsAb). First, as the traditional application of the icIEF method, it serves as an identity assay and purity assay for the BsAb. Second, the method can also be used as an impurity assay for the homodimer monoclonal antibodies generated during BsAb assembly. The homodimer impurity analysis for BsAb is usually done by hydrophobic interaction chromatography methods in the industry. The icIEF method has good sensitivity (down to 4 µg/mL in a limit of quantitation) when UV fluorescence detection is used, which detects the native fluorescence of proteins. This is the first report that an icIEF method has been applied as impurity assay.

Co-chaperonin GroES subunit exchange as dependent on time, pH, protein concentration, and urea.

The discovery of a subunit exchange in some oligomeric proteins, implying short-term dissociation of their oligomeric structure, requires new insights into the role of the quaternary structure in oligomeric protein stability and function. Here we demonstrate the effect of pH, protein concentration, and urea on the efficiency of GroES heptamer (GroES7) subunit exchange. A mixture of equimolar amounts of wild-type (WT) GroES7 and its Ala97Cys mutant modified with iodoacetic acid (97-carboxymethyl cysteine or CMC-GroES7) was incubated in various conditions and subjected to isoelectric focusing (IEF) in polyacrylamide gel. For each sample, there are eight Coomassie-stained electrophoretic bands showing different charges that result from a different number of included mutant subunits, each carrying an additional negative charge. The intensities of these bands serve to analyze the protein subunit exchange. The protein stability is evaluated using the transverse urea gradient gel electrophoresis (TUGGE). At pH 8.0, the intensities of the initial bands corresponding to WT-GroES7 and CMC-GroES7 are decreased with a half-time of (23 ± 2) min. The exchange decreases with decreasing pH and seems to be strongly hindered at pH 5.2 due to the protonation of groups with pK âˆ¼ 6.3, which stabilizes the protein quaternary structure. The destabilization of the protein quaternary structure caused by increased pH, decreased protein concentration, or urea accelerates the GroES subunit exchange. This study allows visualizing the subunit exchange in oligomeric proteins and confirms its direct connection with the stability of the protein quaternary structure.

A high stable sample loading for analysis of adult alpha-thalassemia via the improved microarray isoelectric focusing of Hb species.

The isoelectric focusing has realized various improvements, including the protocols and creation of mIEF (microcolumn isoelectric focusing) instruments with excellent sensitivity for screening of diabetes and beta thalassemia. However, the problem of manual sample loading and hydration for the mIEF limits the operational capacity for stably detecting and quantitating most abnormal hemoglobin (Hb). Herein, we provided a high stable sample loading protocol for analysis of alpha thalassemia and Hb variants. In contrast to the previous volume of 20 µl, a 100 µl blood sample solution in this protocol was optimized with mixture of 6.4-7.5 and 3-10 pH carrier ampholytes, pI markers and loaded for 30 mins IPG microcolumn hydration. The hydrated microcolumn was then automatically loaded onto the mIEF chip array to which CH3COOH and NH4OH act as anodic and cathodic solutions. Lastly, the IEF was run for 9 mins. Hb H, Barts, A1c, F, A2 and CS were simultaneously separated and focused with higher resolution and sensitivity in quantifying H and Barts as low as 0.6 and 0.5 % respectively. Accordingly, there was an enhanced stability and linearity with a rapid assay time of 45 secs per sample. Moreover, analysis showed a fitting linear relationship with conventional technology at R2 = 0.9803 for H and R2 = 0.9728 for Barts thereby indicating greater accuracy confirmed by the AUC. Hence, the developed protocol could simply be employed for high stable and throughput batch sample loading of hydration, and accurate separation and quantitation of Hb variants for alpha and beta thalassemia.

Microstrip isoelectric focusing with deep learning for simultaneous screening of diabetes, anemia, and thalassemia.

BACKGROUND: Hemoglobin (Hb) is an important protein in red blood cells and a crucial diagnostic indicator of diseases, e.g., diabetes, thalassemia, and anemia. However, there is a rare report on methods for the simultaneous screening of diabetes, anemia, and thalassemia. Isoelectric focusing (IEF) is a common separative tool for the separation and analysis of Hb. However, the current analysis of IEF images is time-consuming and cannot be used for simultaneous screening. Therefore, an artificial intelligence (AI) of IEF image recognition is desirable for accurate, sensitive, and low-cost screening. RESULTS: Herein, we proposed a novel comprehensive method based on microstrip isoelectric focusing (mIEF) for detecting the relative content of Hb species. There was a good coincidence between the quantitation of Hb via a conventional automated hematology analyzer and the one via mIEF with R2 = 0.9898. Nevertheless, our results showed that the accuracy of disease diagnosis based on the quantification of Hb species alone is as low as 69.33 %, especially for the simultaneous screening of multiple diseases of diabetes, anemia, alpha-thalassemia, and beta-thalassemia. Therefore, we introduced a ResNet1D-based diagnosis model for the improvement of screening accuracy of multiple diseases. The results showed that the proposed model could achieve a high accuracy of more than 90 % and a good sensitivity of more than 96 % for each disease, indicating the overwhelming advantage of the mIEF method combined with deep learning in contrast to the pure mIEF method. SIGNIFICANCE: Overall, the presented method of mIEF with deep learning enabled, for the first time, the absolute quantitative detection of Hb, relative quantitation of Hb species, and simultaneous screening of diabetes, anemia, alpha-thalassemia, and beta-thalassemia. The AI-based diagnosis assistant system combined with mIEF, we believe, will help doctors and specialists perform fast and precise disease screening in the future.

Monolithically Structured van der Waals Materials for Volume-Polariton Refraction and Focusing.

Polaritons, hybrid light and matter waves, offer a platform for subwavelength on-chip light manipulation. Recent works on planar refraction and focusing of polaritons all rely on heterogeneous components with different refractive indices. A fundamental question, thus, arises whether it is possible to configure two-dimensional monolithic polariton lenses based on a single medium. Here, we design and fabricate a type of monolithic polariton lens by directly sculpting an individual hyperbolic van der Waals crystal. The in-plane polariton focusing through sculptured step-terraces is triggered by geometry-induced symmetry breaking of momentum matching in polariton refractions. We show that the monolithic polariton lenses can be robustly tuned by the rise of van der Waals terraces and their curvatures, achieving a subwavelength focusing resolution down to 10% of the free-space light wavelength. Fusing with transformation optics, monolithic polariton lenses with gradient effective refractive indices, such as Luneburg lenses and Maxwell's fisheye lenses, are expected by sculpting polaritonic structures with gradually varied depths. Our results bear potential in planar subwavelength lenses, integrated optical circuits, and photonic chips.

Multifunctional Meta-Devices for Full-Polarization Rotation and Focusing in the Near-Infrared.

The creation of multi-channel focused beams with arbitrary polarization states and their corresponding optical torques finds effective applications in the field of optical manipulation at the micro-nanoscale. The existing metasurface-based technologies for polarization rotation have made some progress, but they have been limited to single functions and have not yet achieved the generation of full polarization. In this work, we propose a multi-channel and spatial-multiplexing interference strategy for the generation of multi-channel focusing beams with arbitrary polarization rotation based on all-dielectric birefringent metasurfaces via simultaneously regulating the propagation phase and the geometric phase and independently controlling the wavefronts at different circular polarizations. For the proof of concept, we demonstrate highly efficient multi-channel polarization rotation meta-devices. The meta-devices demonstrate ultra-high polarization extinction ratios and high focusing efficiencies at each polarization channel. Our work provides a compact and versatile wavefront-shaping methodology for full-polarization control, paving a new path for planar multifunctional meta-optical devices in optical manipulation at micro-nano dimensions.

Dielectrophoretic separation/classification/focusing of microparticles using electrified lab-on-a-disc platforms.

BACKGROUND: Separation, classification, and focusing of microparticles are essential issues in microfluidic devices that can be implemented in two categories: using labeling and label-free methods. Label-free methods differentiate microparticles based on their inherent properties, including size, density, shape, electrical conductivity/permittivity, and magnetic susceptibility. Dielectrophoresis is an advantageous label-free technique for this objective. Besides, centrifugal microfluidic devices exploit centrifugal forces to move liquid and particles. The simultaneous combination of dielectrophoretic and centrifugal forces exerted on microparticles still needs to be scrutinized more to predict their trajectories in such devices. RESULTS: An integrated system utilizing two categories in microfluidics is proposed: dielectrophoretic manipulation of microparticles and centrifugal-driven microfluidics, followed by a numerical analysis. In this regard, we assumed a rectangular microchannel with internal unilateral planar electrodes equipped with three equal-sized outlets placed radially on a centrifugal platform where microparticles flow toward the disc's outer edge. The effect of different coordinate-based parameters, including radial and lateral distances (X and Y offsets)/tilting angles toward the radius direction (α), on the particles' movement was investigated. Additionally, the effect of operational parameters, including applied voltage, the microchannel width, the number of enabled electrodes, the diameter of particles, and the configuration of electrodes, were analyzed, and the distributions of particles toward the outlets were monitored. It was found that enhanced particle focusing becomes possible at lower rotation speeds and higher electric field values. Furthermore, the proposed centrifugal-DEP system's efficiency for classifying red blood cells/platelets and Live/Dead yeast cells systems was evaluated. SIGNIFICANCE: Our integrated system is introduced as a novel method for focusing and classifying various microparticles with no need for sheath flows, having the ability to conduct particles at desired routes and focusing width. Furthermore, the system effectively separates various bioparticles and offers ease of operation and high-efficiency throughput over conventional dielectrophoretic devices.

Elasto-inertial particle focusing in sinusoidal microfluidic channels.

Dean flow existing in sinusoidal channels could enhance the throughput and efficiency for elasto-inertial particle focusing. However, the fundamental mechanisms of elasto-inertial focusing in sinusoidal channels are still unclear. This work employs four microfluidic devices with symmetric and asymmetric sinusoidal channels to explore the elasto-inertial focusing mechanisms over a wide range of flow rates. The effects of rheological property, flow rate, sinusoidal channel curvature, particle size, and asymmetric geometry on particle focusing performance are investigated. It is intriguing to find that the Dean flow makes a substantial contribution to the particle elasto-inertial focusing. The results illustrate that a better particle focusing performance and a faster focusing process are obtained in the sinusoidal channel with a small curvature radius due to stronger Dean flow. In addition, the particle focusing performance is also related to particle diameter and rheological properties, the larger particles show a better focusing performance than smaller particles, and the smaller flow rate is required for particles to achieve stable focusing at the outlet in the higher concentration of polyvinylpyrrolidone solutions. Our work offers an increased knowledge of the mechanisms of elasto-inertial focusing in sinusoidal channels. Ultimately, these results provide supportive guidelines into the design and development of sinusoidal elasto-inertial microfluidic devices for high-performance focusing.

Probing the Anisotropic Fermi Surface in Tetralayer Graphene via Transverse Magnetic Focusing.

Bernal-stacked tetralayer graphene (4LG) exhibits intriguing low-energy properties, featuring two massive sub-bands and showcasing diverse features of topologically distinct, anisotropic Fermi surfaces, including Lifshitz transitions and trigonal warping. Here, we study the influence of the band structure on electron dynamics within 4LG using transverse magnetic focusing. Our analysis reveals two distinct focusing peaks corresponding to the two sub-bands. Furthermore, we uncover a pronounced dependence of the focusing spectra on crystal orientations, indicative of an anisotropic Fermi surface. Utilizing the semiclassical model, we attribute this orientation-dependent behavior to the trigonal warping of the band structure. This phenomenon leads to variations in electron trajectories based on crystal orientation. Our findings not only enhance our understanding of the dynamics of electrons in 4LG but also offer a promising method for probing anisotropic Fermi surfaces in other materials.

Fast adaptive focusing confocal Raman microscopy for large-area two-dimensional materials.

Large-area two-dimensional (2D) materials possess significant potential in the development of next generation semiconductor due to their unique physicochemical properties. Confocal Raman spectroscopy (CRM), a typical 2D material characterization method, has a limited effective measurement area owing to the restricted focus depth of the system and the less-than-ideal level of the substrate. We propose fast adaptive focusing confocal Raman microscopy (FAFCRM) to realize real-time focusing detection for large-area 2D materials. By observing spot changes on the charge coupled device (CCD) caused by placing an aperture in front of the CCD, the methodology gives a focusing resolution up to 100 nm per 60 µm without axial scanning. A graphene was measured over 25.6 mm × 25.6 mm area on focus through all the scanning. The research results provide new perspectives for non-destructive characterization of 2D materials at the inch level.

Electron wave and quantum optics in graphene.

In the last decade, graphene has become an exciting platform for electron optical experiments, in some aspects superior to conventional two-dimensional electron gases (2DEGs). A major advantage, besides the ultra-large mobilities, is the fine control over the electrostatics, which gives the possibility of realising gap-less and compact p-n interfaces with high precision. The latter host non-trivial states,e.g., snake states in moderate magnetic fields, and serve as building blocks of complex electron interferometers. Thanks to the Dirac spectrum and its non-trivial Berry phase, the internal (valley and sublattice) degrees of freedom, and the possibility to tailor the band structure using proximity effects, such interferometers open up a completely new playground based on novel device architectures. In this review, we introduce the theoretical background of graphene electron optics, fabrication methods used to realise electron-optical devices, and techniques for corresponding numerical simulations. Based on this, we give a comprehensive review of ballistic transport experiments and simple building blocks of electron optical devices both in single and bilayer graphene, highlighting the novel physics that is brought in compared to conventional 2DEGs. After describing the different magnetic field regimes in graphene p-n junctions and nanostructures, we conclude by discussing the state of the art in graphene-based Mach-Zender and Fabry-Perot interferometers.

Microfluidics-assisted fabrication of natural killer cell-laden microgel enhances the therapeutic efficacy for tumor immunotherapy.

Recently, interest in cancer immunotherapy has increased over traditional anti-cancer therapies such as chemotherapy or targeted therapy. Natural killer (NK) cells are part of the immune cell family and essential to tumor immunotherapy as they detect and kill cancer cells. However, the disadvantage of NK cells is that cell culture is difficult. In this study, porous microgels have been fabricated using microfluidic channels to effectively culture NK cells. Microgel fabrication using microfluidics can be mass-produced in a short time and can be made in a uniform size. Microgels consist of photo cross-linkable polymers such as methacrylic gelatin (GelMa) and can be regulated via controlled GelMa concentrations. NK92 cell-laden three-dimensional (3D) microgels increase mRNA expression levels, NK92 cell proliferation, cytokine release, and anti-tumor efficacy, compared with two-dimensional (2D) cultures. In addition, the study confirms that 3D-cultured NK92 cells enhance anti-tumor effects compared with enhancement by 2D-cultured NK92 cells in the K562 leukemia mouse model. Microgels containing healthy NK cells are designed to completely degrade after 5 days allowing NK cells to be released to achieve cell-to-cell interaction with cancer cells. Overall, this microgel system provides a new cell culture platform for the effective culturing of NK cells and a new strategy for developing immune cell therapy.

Imaging of Structural Timber Based on In Situ Radar and Ultrasonic Wave Measurements: A Review of the State-of-the-Art.

With the rapidly growing interest in using structural timber, a need exists to inspect and assess these structures using non-destructive testing (NDT). This review article summarizes NDT methods for wood inspection. After an overview of the most important NDT methods currently used, a detailed review of Ground Penetrating Radar (GPR) and Ultrasonic Testing (UST) is presented. These two techniques can be applied in situ and produce useful visual representations for quantitative assessments and damage detection. With its commercial availability and portability, GPR can help rapidly identify critical features such as moisture, voids, and metal connectors in wood structures. UST, which effectively detects deep cracks, delaminations, and variations in ultrasonic wave velocity related to moisture content, complements GPR's capabilities. The non-destructive nature of both techniques preserves the structural integrity of timber, enabling thorough assessments without compromising integrity and durability. Techniques such as the Synthetic Aperture Focusing Technique (SAFT) and Total Focusing Method (TFM) allow for reconstructing images that an inspector can readily interpret for quantitative assessment. The development of new sensors, instruments, and analysis techniques has continued to improve the application of GPR and UST on wood. However, due to the hon-homogeneous anisotropic properties of this complex material, challenges remain to quantify defects and characterize inclusions reliably and accurately. By integrating advanced imaging algorithms that consider the material's complex properties, combining measurements with simulations, and employing machine learning techniques, the implementation and application of GPR and UST imaging and damage detection for wood structures can be further advanced.

Micropump integrated white blood cell separation platform for detection of chronic granulomatous disease.

White blood cells (WBCs) are robust defenders during antigenic challenges and prime immune cell functioning indicators. High-purity WBC separation is vital for various clinical assays and disease diagnosis. Red blood cells (RBCs) are a major hindrance in WBC separation, constituting 1000 times the WBC population. The study showcases a low-cost micropump integrated microfluidic platform to provide highly purified WBCs for point-of-care testing. An integrated user-friendly microfluidic platform was designed to separate WBCs from finger-prick blood (⁓5 µL), employing an inertial focusing technique. We achieved an efficient WBC separation with 86% WBC purity and 99.99% RBC removal rate in less than 1 min. In addition, the microdevice allows lab-on-chip colorimetric evaluation of chronic granulomatous disease (CGD), a rare genetic disorder affecting globally. The assay duration, straight from separation to disease detection, requires only 20 min. Hence, the proposed microfluidic platform can further be implemented to streamline various clinical procedures involving WBCs in healthcare industries.

Automatic flaw detection in sectoral scans using machine learning.

Phased array ultrasonic testing (PAUT) requires highly trained and qualified personnel to interpret and analyze images. It takes a solid understanding of wave propagation physics to comprehend the generated images. As such, the inspector's judgment and level of experience have a significant impact on the analysis's outcome. In addition, the procedure is prone to error and laborious. AI had shown to be effective in computer vision in a variety of classification and detection tasks. Regarding PAUT, studies have also demonstrated that machine learning may be able to identify defects with a level of accuracy that is on par or even superior to that of trained and qualified inspectors. Nonetheless, the use of computer vision in PAUT remains very limited. The primary cause of this is the challenge accessing large databases of labelled inspections. In fact, a considerable amount of training data is required for machine learning. While it is easy to access sizeable, labelled databases of MRI scans or photographs for instance, that is not the case in PAUT because inspection results are usually confidential. In this project, a large database was generated using mock-ups commonly used to train and evaluate inspectors. The different defects contained in these mock-ups were used to train a machine learning model. The data was acquired with several different probes centered at different frequencies. Each acquisition was performed using Full Matrix Capture (FMC). The post-processing of the data contained in the FMC allows to compute any sectoral scan from its focal laws. As a result, a comprehensive database composed of hundreds of thousands of sectoral scans was generated from these few FMC acquisitions. The completeness of this database facilitated robust training of a defect detection model for PAUT sectoral scans. The evaluation of the model demonstrated its ability to generalize even to defect types it had never been trained on. Furthermore, the detection performance remained consistent even in high noise conditions where the Contrast-to-Noise Ratio (CNR) was very low.

Computational Hyperspectral Microflow Cytometry.

Miniaturized flow cytometry has significant potential for portable applications, such as cell-based diagnostics and the monitoring of therapeutic cell manufacturing, however, the performance of current techniques is often limited by the inability to resolve spectrally-overlapping fluorescence labels. Here, the study presents a computational hyperspectral microflow cytometer (CHC) that enables accurate discrimination of spectrally-overlapping fluorophores labeling single cells. CHC employs a dispersive optical element and an optimization algorithm to detect the full fluorescence emission spectrum from flowing cells, with a high spectral resolution of ≈3 nm in the range from 450 to 650 nm. CHC also includes a dedicated microfluidic device that ensures in-focus imaging through viscoelastic sheathless focusing, thereby enhancing the accuracy and reliability of microflow cytometry analysis. The potential of CHC for analyzing T lymphocyte subpopulations and monitoring changes in cell composition during T cell expansion is demonstrated. Overall, CHC represents a major breakthrough in microflow cytometry and can facilitate its use for immune cell monitoring.

The multi-slit very small angle neutron scattering instrument at the China Spallation Neutron Source.

A multi-slit very small angle neutron scattering (MS-VSANS) instrument has been finally accepted at the China Spallation Neutron Source (CSNS). It is the first spallation neutron source based VSANS instrument. MS-VSANS has a good signal-to-noise ratio and can cover a wide scattering vector magnitude range from 0.00028 to 1.4 Å-1. In its primary flight path, a combined curved multichannel beam bender and sections of rotary exchange drums are installed to minimize the background downstream of the instrument. An exchangeable multi-slit beam focusing system is integrated into the primary flight path, enabling access to a minimum scattering vector magnitude of 0.00028 Å-1. MS-VSANS has three modes, namely conventional SANS, polarizing SANS and VSANS modes. In the SANS mode, three motorized high-efficiency 3He tube detectors inside the detector tank cover scattering angles from 0.12 to 35° simultaneously. In the polarizing SANS mode, a double-V cavity provides highly polarized neutrons and a high-efficiency 3He polarization analyser allows full polarization analysis. In the VSANS mode, an innovative high-resolution gas electron multiplier detector covers scattering angles from 0.016 to 0.447°. The absolute scattering intensities of a selection of standard samples are obtained using the direct-beam technique; the effectiveness of this method is verified by testing the standard samples and comparing the results with those from a benchmark instrument. The MS-VSANS instrument is designed to be flexible and versatile and all the design goals have been achieved.

"Negative" Impact: The Role of Payload Charge in the Physicochemical Stability of Auristatin Antibody-Drug Conjugates.

Antibody-drug conjugates (ADCs) tend to be less stable than their parent antibodies, which is often attributed to the hydrophobic nature of their drug payloads. This study investigated how the payload charge affects ADC stability by comparing two interchain cysteine ADCs that had matched drug-to-antibody ratios and identical linkers but differently charged auristatin payloads, vcMMAE (neutral) and vcMMAF (negative). Both ADCs exhibited higher aggregation than their parent antibody under shaking stress and thermal stress conditions. However, conjugation with vcMMAF increased the aggregation rates to a greater extent than conjugation with uncharged but more hydrophobic vcMMAE. Consistent with the payload logD values, ADC-vcMMAE showed the greatest increase in hydrophobicity but minor changes in charge compared with the parent antibody, as indicated by hydrophobic interaction chromatography and capillary electrophoresis data. In contrast, ADC-vcMMAF showed a decrease in net charge and isoelectric point along with an increase in charge heterogeneity. This charge alteration likely contributed to a reduced electrostatic repulsion and increased surface activity in ADC-vcMMAF, thus affecting its aggregation propensity. These findings suggest that not only the hydrophobicity of the payload, but also its charge should be considered as a critical factor affecting the stability of ADCs.

Two-Dimensional Gel Electrophoresis for Protein Separation of Plant Samples.

This chapter provides a description of the procedure for two-dimensional electrophoresis that can be performed for any given gel size and isoelectric focusing range. This will enable the operator to recognize critical steps and gain sufficient information to generate 2D images suitable for computer-assisted analysis of 2D-gel, as well as mass spectrometry analysis for protein identification and characterization.