Probe Displacement Research Articles

Site-specific detection of circulating tumor DNA methylation in biological samples utilizing phosphorothioated primer-based loop-mediated isothermal amplification

DNA methylation, a kind of epigenetic alteration, plays a vital role in tumorigenesis and offers a new class of targets for cancer treatment. DNA hypermethylation at the E-Box site (CACGTG, −288 bp) in the SLC22A2 promoter was related to multidrug resistance of renal cell carcinoma (RCC), which can provide the target for both treatment and monitoring. Herein, we developed a novel phosphorothioated primer based loop-mediated isothermal amplification (PS-LAMP) assay to detect circulating tumor DNA (ctDNA) methylation levels in E-Box sites in tumor tissue, urine, and plasma samples from patients with RCC. Bisulfite treatment converted methylated/unmethylated discrepancy to a single base discrepancy (C/U). PS-LAMP amplified the templates to a tremendous amount. One-step strand displacement (OSD) probe provided single base resolution in amplified products and finally realized the specific site methylation detection. Our proposed method provided a linear range from 0% to 100% for methylation levels and was available in samples at low concentrations (102 copies/μL). Visually observable colorimetric detection can be achieved by incorporating the OSD probe with gold nanoparticles (AuNP). Our assay performed better than traditional methods in biological samples with low ctDNA concentration. Further, we found a potential consistency of methylation levels between tumor tissue and plasma sample from the same patient (Spearman's ρ = 0.886, P = 0.019, n = 6). In general, this work provides a PS-LAMP assay combining OSD probes for site-specific methylation detection in various biological samples, offering a method for noninvasive detection.

Transcranial Doppler Remote Positioning System with Virtual Reality Integration for Vestibular Studies.

Transcranial doppler (TCD) ultrasound probes are an invaluable tool in cerebral blood flow (CBF) studies. Their operation demands maintaining consistent pose on the subject throughout the experimental protocol. However, the displacement of the TCD probe during vestibular studies is common and substantially prolongs the experiment or even terminates it. This is a significant challenge for integrating motion-based vestibular studies with CBF investigations. In response, a mechatronics system is designed to allow remote repositioning of the TCD probe during data collection experiments while the subject is wearing a head mounted virtual reality (VR) display and seated in a vestibular disorientation device. This paper presents the design, prototype, and operation of this mechatronics apparatus.Clinical Relevance- The mechatronics apparatus of this paper can enable motion-based vestibular studies that entail the use of CBF velocity measurement and head-mounted virtual reality display.

Analysis of macerated ticks using Boolean logic gating colorimetric isothermal nucleic acid assays for Lyme Borrelia and Ixodes scapularis ticks

Lyme disease, one of the most common tickborne diseases, has been rapidly spreading in parallel with the expansion of the range of its tick vector. Better tick surveillance efforts are needed to accurately estimate disease risk and to guide public health and clinical management. We have developed two multiplex loop-mediated isothermal amplification (LAMP) reactions coupled with oligonucleotide strand displacement (OSD) probes to identify the tick host, Ixodes scapularis, and the Lyme disease pathogen, Borrelia burgdorferi, they carry. In each multiplex LAMP-OSD assay the co-presence of two target sequences is computed at the DNA level by linking the two corresponding amplicons and detecting the co-product on colorimetric lateral flow dipsticks. In tests with synthetic DNA, the co-presence of as few as four copies of input DNA could be detected, without producing spurious signals. Most importantly, though, the LAMP-OSD assay is amenable to being carried out directly with macerated tick samples, without any sample preparation. In such field conditions, assays performed robustly and demonstrated 97–100% sensitivity and 100% specificity with both field-collected and lab-raised artificially infected ticks. Such easy-to-use, arthropod and pathogen-specific assays would be well suited to field and near patient use without relying on complex instrumentation or infrastructure.

Reaction–Diffusion Process for Hydrogels with a Tailored Layer Structure

The architecture of hydrogel composites results in not only synergistic property enhancement but also superior functionality. The reaction–diffusion (RD) process is acommon phenomenon throughout nature which induced ordered structure on a length scale from microscopic to macroscopic. Different from commonly used inorganic salts or oppositely charged nanoparticles for the RD process, a modified RD process was used for layered chitosan hydrogel (L-CH) and layered magnetic chitosan hydrogel (L-MCH). During the modified RD process reported in this paper, the protonated chitosan (CS-NH3+) with iron ions (Fe3+ and Fe2+) was used as an inner-reactant and hydroxide ion (-OH−) was used as an out-reactant. The protonated chitosan (CS-NH3+) not only played the role of an inner-reactant but also the reaction medium which controlled the diffusion behavior of the out-reactant (-OH−). A series of ordered layers were constructed and the ordered layers were parallel with the longitudinal axis. The layer width of L-CH and L-MCH can be tailored by varying interval time T. The mean layer width of L-MCH increased from 50 ± 5.8 μm to 90 ± 6.4 μm when the interval time T increased from 2 min to 5 min. The tailored layer structure of L-CH and L-MCH obeyed the time law and spacing law, which declared that the L-CH and L-MCH were constructed via the reaction–diffusion process. We also show that the tailored layer structure endows hydrogel with enhanced mechanical properties, especially toughness. The yield strength of magnetic chitosan hydrogel was improved significantly (from 95.1 ± 7.6 kPa to 401.7 ± 12.1 kPa, improved by about 4 times) when 10 wt. % magnetite nanoparticles were involved. The enhancement of the mechanical propertieswas due to a physical crosslinking effect of magnetite nanoparticles on chitosan. For L-MCH, the probe displacement reached 28.93 ± 2.6 mm when the rupture occurred, which was as high as 284.7% compared with that of the non-layered hydrogel. The tailor-made hydrogels might be possible for application as a tough implantable scaffold.

In-vivo Comparison of Planned and Measured Rectal Doses during Cobalt-60 HDR CT-based Intracavitary Brachytherapy Applications of Cervical Cancer Using the PTW 9112 Semiconductor Probe.

A semiconductor rectal probe was used to compare planned and measured rectal doses during Co-60 high dose rate (HDR) CT-based intracavitary brachytherapy applications (ICBT) of cervical cancer. A total of 22 HDR brachytherapy applications were included from 11 patients who were first treated with EBRT to the whole pelvis with a total prescribed dose of 50 Gy in 25 fractions. During each application, a PTW 9112 probe rectal probe having a series of five semiconductor diodes (R1 to R5) was inserted into the patient's rectum and a CT-based HDR ICBT application with a prescribed dose per fraction of 7 or 7.5 Gy to HRCTV was performed. Measurements were carried in water phantom using PTW rectal and universal adaptor plugs. Doses measured in phantom and with patients were compared to those calculated by the treatment planning system. The mean percentage dose difference ΔD (%) between calculated and measured values from phantom study were -5.29%, 1.89%, -2.72%, -4.76, and 0.72% for R1, R2, R3, R4, and R3 diodes, respectively and the overall mean ΔD (%) value with standard deviation (SD) was -2.03%±9.6%. From the patient study, a ΔD (%) that ranged from -19.5% to 24.0%, which corresponded to dose disparities between -0.77 Gy and 0.66 Gy. The median ΔD (%) ranged from 0.4% to 1.3%, or -0.03 to 0.05 Gy, respectively. ΔD (%) values exceeded 10% in approximately 26.4% of measurements (29 out of 110 in 22 applications). The location of Rmax in computed and measured values differs in 5 of 22 applications might be due to possible displacement of rectal probe between simulation and treatment. Despite the likely geometrical shift of measuring detectors between insertion and treatment, in-vivo dosimetry is feasible and can be used to estimate the dose to the rectum during HDR ICBT.

Recent advances toward mesoscopic quantum optomechanics

We present a number of approaches, currently in experimental development in our research groups, toward the general problem of macroscopic quantum mechanics, i.e., manifestations of quantum noise and quantum fluctations with macroscopic (engineered and microfabricated by man) mechanical systems. Discussed experiments include a pendulum, a torsion pendulum, a ng-scale phononic-crystal silicon nitride membrane, a μ g-scale quartz resonator, and mg-scale mirrors for optical levitation. We also discuss relevant applications to quantum thermometry with optomechanical systems and the use of squeezed light to probe displacements beyond conventional quantum limits.

Investigations of In-Plane Fluidelastic Instability in a Multispan U-Bend Tube Array—Part II: Tests in Two-Phase Flow

Abstract Tests to study fluidelastic instability (FEI) in an array of U-bend tubes were recently completed in the multispan U-Bend test rig at Canadian Nuclear Laboratories. These tests were sponsored by the Electric Power Research Institute and were designed to study in-plane fluid elastic instability of steam generator tubes in two-phase cross flow. This instability mechanism was first observed in previous experiments by Atomic Energy of Canada Limited. This mechanism was not thought to be a serious practical concern until 2012 when it caused severe damage to tubes in a new replacement steam-generator in a nuclear power plant in the United States. In this study, tests were conducted both with flows of air and two-phase liquid/vapor Refrigerant 134a. The tube bundle consisted of 22 flexible U-bend tubes supported by a configurable flat-bar arrangement. Testing focused on the effects of support geometry and tube-to-support interaction. Data were recorded from 33 dynamic signals from accelerometers, displacement probes, force transducers, and void-fraction probes. Part I of this two-part series presented results of air tests. Part II presents results of tests using two-phase Freon refrigerant (R-134a) as the working fluid.

A novel DNA detection using spherical identification probe and strand displacement reaction-initiated silver nanocluster switch.

Herein, we developed a novel fluorescent assay using spherical identification probes and toehold-mediated strand displacement reaction-initiated silver nanoclusters (AgNCs) "on-off" signal switch. In this strategy, the target was captured by the spherical probes to induce the activity of exonuclease III (Exo III), catalyzing the cyclic cleavage of substrates to produce a mass of trigger strands. After magnetic bead separation, the intermediates in the supernatant activated downstream toehold-mediated strand displacement reaction to change the structure of silver nanocluster templates, leading to fluorescence intensity reduction. Furthermore, it is demonstrated that the application of spherical identification probes could reduce the signal leakage and the limit of detection. In addition, AgNCs with perfect optical property were ingeniously combined to realize signal output, which reduced the cost and time of synthesis. Under the optimal conditions, the sensing method displayed a good linear range from 250pM to 25nM with a detectable minimum concentration of 250pM. And the practical application potential in complex biological matrices was also evaluated. Considering these advantages, this constructed strategy opens a new path for nucleic acid detection with better performance. A simple, label- and hairpin-free fluorescent system based on spherical identification probe and toehold-mediated strand displacement reaction-initiated silver nanoclusters (AgNCs) "on-off" signal switch was successfully constructed to detect target DNA.

Fluorine-thiol displacement probes for acetaminophen's hepatotoxicity.

Chemicals possessing reactive electrophiles can denature innate proteins leading to undesired toxicity, and the overdose-induced liver injury by drugs containing electrophiles has been one of the major causes of non-approval and withdraw by the US Food and Drug Administration (FDA). Elucidating the associated proteins could guide the future development of therapeutics to circumvent these drugs' toxicities, but was largely limited by the current probing tools due to the steric hindrance of chemical tags including the common "click chemistry" labels. Taking the widely used non-steroidal anti-inflammatory drug acetaminophen (APAP) as an example, we hereby designed and synthesized an APAP analogue using fluorine as a steric-free label. Cell toxicity studies indicated our analogue has similar activity to the parent drug. This analogue was applied to the mouse hepatocellular proteome together with the corresponding desthiobiotin-SH probe for subsequent fluorine-thiol displacement reactions (FTDRs). This set of probes has enabled the labeling and pull-down of hepatocellular target proteins of the APAP metabolite as validated by Western blotting. Our preliminary validation results supported the interaction of APAP with the thioredoxin protein, which is an important redox protein for normal liver function. These results demonstrated that our probes confer minimal steric perturbation and mimic the compounds of interest, allowing for global profiling of interacting proteins. The fluorine-thiol displacement probing system could emerge as a powerful tool to enable the investigation of drug-protein interactions in complex biological environments.

A novel linear displacement isothermal amplification with strand displacement probes (LDIA-SD) in a pocket-size device for point-of-care testing of infectious diseases

Nucleic acid amplification is crucial for disease diagnosis, especially lethal infectious diseases such as COVID-19. Compared with PCR, isothermal amplification methods are advantageous for point-of-care testing (POCT). However, complicated primer design limits their application in detecting some short targets or sequences with abnormal GC content. Herein, we developed a novel linear displacement isothermal amplification (LDIA) method using two pairs of conventional primers and Bacillus stearothermophilus (Bst) DNA polymerase, and reactions could be accelerated by adding an extra primer. Pseudorabies virus gE (high GC content) and Salmonella fimW (low GC content) genes were used to evaluate the LDIA assay. Using strand displacement (SD) probes, a LDIA-SD method was developed to realize probe-based specific detection. Additionally, we incorporated a nucleic acid-free extraction step and a pocket-sized device to realize POCT applications of the LDIA-SD method. The LDIA-SD method has advantages including facile primer design, high sensitivity and specificity, and applicability for POCT, especially for amplification of complex sequences and detection of infectious diseases.

A Fiber-Based Chromatic Dispersion Probe for Simultaneous Measurement of Dual-Axis Absolute and Relative Displacement.

This paper presents a fiber-based chromatic dispersion probe for the simultaneous measurement of dual-axis absolute and relative displacement with nanometric resolutions. The proposed chromatic dispersion probe is based on optical dispersion. In the probe, the employed light beam is split into two sub-beams, and then the two sub-beams are made to pass through two optical paths with different optical settings where two identical single-mode fiber detectors are located at different defocused positions of the respective dispersive lenses. In this way, two spectral signals can be obtained to indicate the absolute displacement of each of the dual-axes. A signal processing algorithm is proposed to generate a normalized output wavelength that indicates the relative displacement of the dual-axis. With the proposed chromatic dispersion probe, the absolute and relative displacement measurements of the dual-axis can be realized simultaneously. Theoretical and experimental investigations reveal that the developed chromatic dispersion probe realizes an absolute measurement range and a measurement resolution of approximately 180 μm and 50 nm, respectively, for each axis. Moreover, a relative displacement measurement range and a measurement resolution of about 240 μm and 100 nm, respectively, are achieved for the dual-axis.

Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification

• We developed a novel method for pathogen detection in foods. • It combined QBs-labelled LFIAS and SD-LAMP to achieve quantitative detection. • The combined test rapidly and sensitively detected Salmonella Thyphimurium. • Current approach does not require almost any equipment allowing point-of-care use. • The efficacy of the current test was confirmed in different food samples. Rapid, sensitive, point-of-care detection of pathogenic bacteria is important for food safety. In this study, we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip (QBs-labelled LFIAS) combined with strand displacement loop-mediated isothermal amplification (SD-LAMP) for quantitative Salmonella Typhimurium (ST) detection. Quantum dot nanobeads (QBs) served as fluorescence reporters, providing good detection efficiency. The customizable strand displacement (SD) probe was used in LAMP to improve the specificity of the method and prevent by-product capture. Detection was based on a sandwich immunoassay. A fluorescence strip reader measured the fluorescence intensity (FI) of the test (T) line and control (C) line. The linear detection range of the strip was 10 2 –10 8 CFU·mL −1 . The visual limit of detection was 10 3 CFU·mL −1 , indicating that the system was 10-fold more sensitive than AuNPs-labelled test strips. ST specificity was analyzed in accordance with agarose gel outputs of PCR and SD-LAMP. We detected ST in foods with an acceptable recovery of 85%–110%. The method is rapid, simple, almost equipment-free, and suitable for bacterial detection in foods and for clinical diagnosis.

Spiral scanning nano-profiler using normal vector tracing method.

Unlike interferometers that obtain data from the entire sample, three-dimensional (3D) measuring instruments acquire data from multiple points. A nano-profiler measures slopes at different points on the surface to determine the sample shape, while a coordinate measuring machine (CMM) utilizes contact or non-contact height displacement probes for taking measurements. Therefore, the 3D measurement time increases if the area to be measured is wide. This leads to a reduced measurement accuracy owing to environmental changes. To reduce the 3D measurement time, a nano-profiler that rotates measured objects at a constant velocity and quickly scans each measurement point by following a spiral trajectory was designed. The developed nano-profiler measures angular distribution instead of height distribution. This makes it possible to reduce the drive error during rotation. Compared to an interferometer, the measurement time was reduced from 163 to 17minwith the nano-profiler and a 10nm peak-to-valley difference was achieved. The slope measurement with spiral scanning was confirmed to be a quick and accurate shape measurement technique that can be used while manufacturing high-precision optical components.

Radial Error Motion Measurement and Its Uncertainty Estimation of Ultra Precision Axes of Rotation with Nanometer Level Precision.

The radial error is the key performance indicator of ultra-precision axis. In order to measure and evaluate the radial error of ultra-precision axis with nanometer accuracy, a measurement system with an accuracy of nanometer based on capacitive displacement probes and standard spheres is developed. The nonlinearity error of capacitive displacement probes, misalignment error of the probes, eccentric error of standard spheres, error caused by environment temperature change, environment vibration and error separation methods are analyzed and the effects of the above factors are obtained; Multiple measurement examples carried out with the measurement system this paaper constructed indicate the repeatability of the measurement system reaches 10.5 nm and the roundness error of artifact separated is less than 4.03 nm. In order to evaluate the measurement dispersion of the ultra-precision axis radial error, the major uncertainty components and the complete process of the comprehensive evaluation of the measurement uncertainty are proposed. The combined uncertainty of radial error motion measurement of the ultra-precision axis with Donaldson reversal is 31.64 nm (k = 2).

Investigation on small molecule-aptamer dissociation equilibria based on antisense displacement probe

Food safety is a major issue to public health and have attracted global attention. Fast, sensitive, and reliable detection methods for food hazardous substances is highly desirable. Aptamers which can bind to the target molecules with high affinity and specificity represent an attractive tool for the recognition of food hazardous substances, which play an important role in the development and application of new food safety detection technology. But current assays for characterizing small molecule-aptamer binding are limited by either the mass sensitivity or the size differentiation ability. Herein, we proposed a comprehensive method for assessing the dissociation equilibria of small molecule-aptamer, which is immobilized-free under ambient conditions. The design employs the Le Chatelier's principle and could be used to effectively measure small molecule-aptamer interactions. ATP binding aptamer and anti-aflatoxin B1 aptamer were used as the model system to determine their affinity, in which their dissociation equilibria measurements are in excellent close to their previous work. Due to the simplicity and sensitivity of this new method, we believe that it could be recommended as an effective tool for characterizing small molecule-aptamer interactions and promote the further application of small molecular aptamer in food safety.

A new three-dimensional elastography using phase based shifted Fourier transform

Elastography is an imaging technique with the ability to determine low quantities of some of the mechanical properties of tissues. The aim of our research is to design a new 3D algorithm using the Shifted Fourier Transform (SFT) to perform a quasi-static elastography. Our innovative idea is implementation of a 3D convolution instead of using three 2D convulsions. At first, we collected the raw data from Abaqus engineering software in the form of breast tissue with a coefficient of elasticity of healthy tissue and tumor tissue with a coefficient of elasticity of tumor tissue. The primary raw data consists of a number of points with x, y and z specified for tumor and healthy breast tissue. At this step, we simulated the displacements in directions of x, y and z at each point of the prescribed tissues for 15 ​mm displacement of probe in –Y direction then we collected 1831 points for tumor and 4186 points for breast before and after pressure. After applying a novel reconstruction algorithm, we convolved all images with the 3D Gabor filters to obtain phases, represented displacements of the breast and tumor images for before and after pressure. To reach this goal, we designed a Gabor filter bank based on the dimensions of the input images in different scales, directions, and deviations. Using the 3D SFT, we calculated the displacements of the breast and tumor tissues followed by 3D elastogram representation of the images. Finally, we implemented a 2D analysis of SFT in order to investigate validation of the 3D SFT. In 2D algorithm, we used three two-dimensional convulsions in XY, YZ and XZ planes. The results obtained from the small displacements marked by circles, confirmed the accuracy of the 3D SFT algorithm. These areas of interest are the tumor areas in the 2D analysis.

Testing beat perception without sensory cues to the beat: the Beat-Drop Alignment Test (BDAT)

Beat perception can serve as a window into internal time-keeping mechanisms, auditory–motor interactions, and aspects of cognition. One aspect of beat perception is the covert continuation of an internal pulse. Of the several popular tests of beat perception, none provide a satisfying test of this faculty of covert continuation. The current study proposes a new beat-perception test focused on covert pulse continuation: The Beat-Drop Alignment Test (BDAT). In this test, participants must identify the beat in musical excerpts and then judge whether a single probe falls on or off the beat. The probe occurs during a short break in the rhythmic components of the music when no rhythmic events are present, forcing participants to judge beat alignment relative to an internal pulse maintained in the absence of local acoustic timing cues. Here, we present two large (N > 100) tests of the BDAT. In the first, we explore the effect of test item parameters (e.g., probe displacement) on performance. In the second, we correlate scores on an adaptive version of the BDAT with the computerized adaptive Beat Alignment Test (CA-BAT) scores and indices of musical experience. Musical experience indices outperform CA-BAT score as a predictor of BDAT score, suggesting that the BDAT measures a distinct aspect of beat perception that is more experience-dependent and may draw on cognitive resources such as working memory and musical imagery differently than the BAT. The BDAT may prove useful in future behavioral and neural research on beat perception, and all stimuli and code are freely available for download.

Inhibitory Effects of Myriocin on Non-Enzymatic Glycation of Bovine Serum Albumin.

Advanced glycation end products (AGEs) are the compounds produced by non-enzymatic glycation of proteins, which are involved in diabetic-related complications. To investigate the potential anti-glycation activity of Myriocin (Myr), a fungal metabolite of Cordyceps, the effect of Myr on the formation of AGEs resulted from the glycation of bovine serum albumin (BSA) and the interaction between Myr and BSA were studied by multiple spectroscopic techniques and computational simulations. We found that Myr inhibited the formation of AGEs at the end stage of glycation reaction and exhibited strong anti-fibrillation activity. Spectroscopic analysis revealed that Myr quenched the fluorescence of BSA in a static process, with the possible formation of a complex (approximate molar ratio of 1:1). The binding between BSA and Myr mainly depended on van der Waals interaction, hydrophobic interactions and hydrogen bond. The synchronous fluorescence and UV-visible (UV-vis) spectra results indicated that the conformation of BSA altered in the presence of Myr. The fluorescent probe displacement experiments and molecular docking suggested that Myr primarily bound to binding site 1 (subdomain IIA) of BSA. These findings demonstrate that Myr is a potential anti-glycation agent and provide a theoretical basis for the further functional research of Myr in the prevention and treatment of AGEs-related diseases.

A Double FBGs Temperature Self-Compensating Displacement Sensor and Its Application in Subway Monitoring.

In order to ensure the effective vibration–reduction and vibration–isolation of the steel spring floating plate rail and meet the safe operation requirements of the subway, a Fiber Bragg Grating (FBG) displacement sensor for the deformation monitoring of the subway floating plate is proposed. The sensor adopts double FBGs to realize temperature self-compensation. The elastic ring is used as the elastic conversion structure after the fiber grating is pre-stretched; the two ends are pasted and fixed in the groove in the diameter direction of the ring, which avoids the waveform distortion caused by the full pasting of the fiber grating. The combination of linear bearing and displacement probe rods can increase stability and reduce friction loss so that the sensor has the advantages of high sensitivity and accurate measurement results. The test results and error analysis show that in the range of 0~20 mm, the sensitivity of the sensor is 164.2 pm/mm, the accuracy reaches 0.09% F.S, and the repeatability error and hysteresis error are only 1.86% and 0.99%, respectively. The thermal displacement coupling experiment proves that the sensor has good temperature self-compensation performance. It provides a new technical scheme for the effective monitoring and condition assessment of the built-in steel spring floating plate rail.

Nanofabrication mechanism of [formula omitted]/20 features achieved by coupling field tip enhancement induced with nanosecond laser irradiating AFM probe tip

The nanofabrication mechanism of coupling field tip enhancement induced by laser-irradiated probe tip was investigated theoretically and experimentally. The coupling mechanisms and distribution characteristics of the enhanced electric, thermal, and stress fields were studied at wavelengths of 400–800 nm and tip-sample distances of 1–20 nm for 9 material combinations. The simulation results demonstrated that the electro-thermal coupling effect and thermal–mechanical coupling effect are the basic principles of this technique, independent of the materials. It was also demonstrated that probe damage due to mechanical scratching can be avoided through the estimation of probe displacement. The superposition of electromagnetic energy and thermal energy generates an extremely constrained total energy of up to 10–12 orders of magnitude near the tip end. The nano-line structures were processed at each tip-sample distance, and nanostructures with a feature size of approximately λ/20 were obtained. The experimental results and the total energy distribution demonstrated that the interaction of the coupling field tip enhancement with samples is in the form of total energy deposits on materials. The significance of this study will not only enable a refined understanding of the fundamental physics of surface nanostructuring but will also advance the scientific and industrial applications of the technology.