Microscopy Technology Research Articles

Structure identification and activity evaluation of polysaccharide from Codonopsis pilosula (C. pilosula nannf. Var. modesta (nannf.) L. T. Shen)

Codonopsis pilosula is a medicinal plant with properties related to food, and the antioxidant and hypoglycemic components of its natural polysaccharides, as well as their related mechanisms, have not been fully understood. This study examined the physicochemical properties and structure of C. pilosula polysaccharide (WCP) using an orthogonal extraction method involving hot water-alcohol precipitation, high-resolution spectroscopy, and microscopic imaging technology. The antioxidant and hypoglycemic activities of the polysaccharides were assessed. Six fractions (WCP1, WCP2, WCP3, WCP4, WCP5, and WCP6) were identified through purification using a DEAE-cellulose column and Sephadex G-100 gel column. Mannose, glucose, and arabinose were identified as the primary monosaccharide components of WCPs through Gas Chromatography (GC). The polysaccharide exhibited a crystal structure composed of irregularly entangled pyranose particles of WCPs as revealed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The polysaccharide decomposed into smoother triple-helix fibrous flocs as indicated by Congo red analysis and circular dichroism analysis (CD). The antioxidant capacity of the polysaccharides against various free radicals was evaluated, with WCP4 demonstrating a significant reducing capacity. Moreover, WCP3 and WCP5 displayed notable inhibition rates of α-amylase and α-glucosidase, respectively. Molecular docking studies indicated the interaction of WCP5 with two enzymes, forming multiple hydrogen bonds through specific amino acids. In the gromacs simulation, both composite systems maintained a steady state in terms of root mean square deviation (RMSD), root mean square fluctuation (RMSF), and radius of gyration, providing strong support for their potential as a reference for the developing novel therapeutic approaches to diabetes.

ClearScope: a fully integrated light sheet theta microscope for sub-cellular resolution imaging without lateral size constraints.

Three-dimensional (3D) ex vivo imaging of cleared intact brains of animal models and large human and non-human primate postmortem brain specimens is important for understanding the physiological neural network connectivity patterns and the pathological alterations underlying neuropsychiatric and neurological disorders. Light-sheet microscopy has emerged as a highly effective imaging modality for rapid high-resolution imaging of large cleared samples. However, the orthogonal arrangements of illumination and detection optics in light sheet microscopy limits the size of specimen that can be imaged. Recently developed light sheet theta microscopy (LSTM) technology addressed this by utilizing a unique arrangement of two illumination light paths oblique to the detection light path, while allowing perpendicular arrangement of the detection light path relative to the specimen surface. Here, we report development of a next-generation, fully integrated, and user-friendly LSTM system for rapid sub-cellular resolution imaging uniformly throughout a large specimen without constraining the lateral (XY) size. In addition, we provide a seamlessly integrated workflow for image acquisition, data storage, pre- and post-processing, enhancement, and quantitative analysis. We demonstrate the system performance by high-resolution 3D imaging of intact mouse brains and human brain samples, and complete data analysis including digital neuron tracing, vessel reconstruction and design-based stereological analysis in 3D. This technically enhanced and user-friendly LSTM implementation will enable rapid quantitative mapping of molecular and cellular features of interests in diverse types of very large samples.

Intradermal Delivery of Calcium Hydroxylapatite With Fractionated Ablation.

The absorption of biostimulatory particulate matter following its application to fractional skin defects remains poorly understood, and even less is known about its in vivo impact in terms of tissue integration. The objectives of this study are twofold: (1) to evaluate the potential of calcium hydroxylapatite (CaHA) to penetrate through skin treated with a fractional laser; and (2) to assess the effectiveness of clinical laser scanning microscopy technologies in monitoring the effects of such treatment over time. One area on a volunteer's arm was treated with a fractional erbium laser (Sciton Inc., Palo Alto, CA), while a second area received the same laser treatment followed by CaHA topical application. We used reflectance confocal microscopy (RCM) and multiphoton microscopy (MPM) to noninvasively image beneath the surface of the treated skin to study and monitor the effects of these treatments within 1 h of treatment and at four additional time points over a 6-week period. One hour posttreatment, at different depths beneath the skin surface, MPM and RCM provided similar visualizations of laser-induced channels. In skin treated by both laser and CaHA, these two imaging methods provided complementary information. RCM captured the lateral and depth distribution of CaHA microspheres and were seen as bright spheres as they became incorporated into the healing tissue. MPM, meanwhile, visualized the CaHA microparticles as dark shadow spheres within the laser-induced channels and encroaching healing tissue. Furthermore, MPM provided critical information about collagen regeneration around the microspheres, with the collagen visually marked by its distinct second harmonic generation (SHG) signal. This observational pilot study demonstrates that CaHA, a collagen stimulator used as a dermal filler, can not only be inserted into the dermis after fractional laser treatment but remains in the healing skin for at least 6 weeks posttreatment. The noninvasive imaging techniques RCM and MPM successfully captured the presence of CaHA microspheres mid-dermis during the healing phase. They also demonstrated new collagen production around the microspheres, highlighting the effectiveness of these imaging approaches in monitoring such treatment over time.

Intraoperative margin assessment during radical prostatectomy: is microscopy frozen in time or ready for digital defrost?

Intraoperative frozen section (IFS) is used with the intention to improve functional and oncological outcomes for patients undergoing radical prostatectomy (RP). High resource requirements of IFS techniques such as NeuroSAFE may preclude widespread adoption, even if there are benefits to patients. Recent advances in fresh-tissue microscopic digital imaging technologies may offer an attractive alternative, and there is a growing body of evidence regarding these technologies. In this narrative review, we discuss some of the familiar limitations of IFS and compare these to the attractive counterpoints of modern digital imaging technologies such as the speed and ease of image generation, the locality of equipment within (or near) the operating room, the ability to maintain tissue integrity, and digital transfer of images. Confocal laser microscopy (CLM) is the modality most frequently reported in the literature for margin assessment during RP. We discuss several imitations and obstacles to widespread dissemination of digital imaging technologies. Among these, we consider how the 'en-face' margin perspective will challenge urologists and pathologists to understand afresh the meaning of positive margin significance. As a part of this, discussions on how to describe, categorize, react to, and evaluate these technologies are needed to improve patient outcomes. Limitations of this review include its narrative structure and that the evidence base in this field is relatively immature but developing at pace.

Effect of Grease Composition on Impact-Sliding Wear

Impact-sliding experiments were performed by using four self-made lithium-based greases, namely Yangtze Grease 1, Yangtze Grease 2, Yangtze Grease 3, and Yangtze Grease 4. The influence of base oil viscosity, thickener content, and morphology of thickener fiber clusters on the lubricating state were visually explored, combined with field-emission microscopy and two-light interference technology. The grease film distribution at the middle section was measured using Dichromatic Interference Intensity Modulation (DIIM) software. All experiments were executed in a completely flooded environment. The results show that among the components of grease, the base oil’s viscosity has the greatest impact on the anti-wear performance of the grease. As the viscosity of the base oil increases, the grease exhibits better anti-wear performance. The grease film thickness under the condition of high-viscosity base oil is about 10 times higher than that under the condition of low-viscosity base oil. Secondly, the content of thickener in the grease needs to be controlled within a reasonable range. The experiments indicate that the effect of thickener content on the grease’s film-forming properties becomes more pronounced at higher speeds. From the experiment using YG 4, it can be seen that a higher thickener content under high-speed conditions increases the thickness of the lubricating grease film by about 10 times. The dimensions of the thickener fibers and the density of their entanglement structure significantly influence the rheological properties and load-bearing capacity of the grease. Larger fiber sizes and higher entanglement densities result in reduced grease fluidity and recovery but enhance its load-bearing capabilities. In order to obtain the best anti-wear performance during impact-sliding motion, the size of the thickener fiber and the density of the entanglement structure need to be controlled within an appropriate range.

Pretreatment with nano-silver extends the post-harvest longevity of gladiolus cut flowers by reducing free water mobility

The water content of cut flowers is a significant factor in their post-harvest quality. In this study, we examine the efficacy of silver nanoparticles (NS) on the longevity of cut gladiolus, with a focus on water state and distribution. We used Low-field nuclear magnetic resonance (LF-NMR) technology to identify three water fractions with different transverse relaxation times (T2) values: bound water T21 (<10 ms), intermediate immobilized water T22 (10–100 ms), and the slowest component free water T23 (>10 ms). During the opening process, T23 increased at stages 2 and 3 and then decreased, T22 decreased slowly, and T21 remained unchanged. Free water values were consistently higher than bound water and immobilized water and reached their maximum from stage 2 until stage 4, when the petals were extended and began to wilt. The vascular bundles responsible for transporting water had higher water content, as detected by proton density-weighted magnetic resonance imaging (MRI). Bound water and free water with NS pretreatments in bracts were initially lower but then two days later the signal amplitude of each water state exceeded those of the control, indicating that the treatment enhanced the water-holding capacity over time. Furthermore, NS pretreatments reduced the free water mobility of the cut flowers and inhibited stem decay. Additionally, we found that NS can enter the stem and are primarily transported upward along the xylem with water using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) technology. Overall, our findings indicate that NS pretreatment reduces free water in gladiolus cut flowers, enhancing their water retention and prolonging their vase life.

Diagnostic Strategies in the Era of Monkeypox Resurgence: A Comprehensive Analysis.

The resurgence of monkeypox (Mpox), an orthopoxvirus infection closely related to smallpox, presents a significant global health challenge. This study presents a comprehensive overview of Mpox, focusing on its clinical manifestations, diagnostic strategies, and testing methodologies. A thorough review of the literature and available data on Mpox, emphasizing diagnostic assays, clinical indicators, and laboratory testing, constitutes the core of this analysis. The study involves insights from Mpox patients and healthcare professionals engaged in its diagnosis and management. Contextualizing the research within the global spread of Mpox addresses the complexities associated with the diagnosis of the disease. The findings illuminate diverse Mpox diagnostic techniques, encompassing viral culture, immunological methods, serology,quantitative polymerase chain reaction (qPCR), electron microscopy, and advanced technologies such as artificial intelligence (AI) and the GeneXpert system. qPCR is highlighted as the benchmark for MPXV detection and quantification. These diagnostic advancements have significantly enhanced the precision and efficiency of Mpox diagnosis, facilitating prompt identification and treatment of infected individuals. The study underscores the critical importance of accurate and timely diagnosis, proper handling and transportation of clinical specimens, and the imperative for point-of-care (POC) testing to control the global spread of Mpox.

Revealing the Molecular Interaction between CTL Base Oil and Additives and Its Application in the Development of Gasoline Engine Oil

In order to improve fuel economy to meet the standard for passenger car oil, a new formulation with good viscosity–temperature performance for gasoline engine oil is required. In this study, coal-to-liquid (CTL) base oil, with a high viscosity index and good low-temperature performance, was selected as the base oil to develop the gasoline engine oil. A systematic study on the molecular interaction between the CTL base oil and the viscosity index improver (VII), including three kinds of hydrogenated styrene diene copolymers (HSD-type) and four kinds of ethylene propylene copolymers (OCP-type), was conducted. It was found that in general, in CTL base oil, the HSD-type VII exhibited a much higher viscosity index, a significantly lower shear stability index, a higher thickening ability, and a lower cold-cranking simulator (CCS) viscosity than that of OCP-type VII. Moreover, when comparing CTL base oil with mineral oil 150N, the combination of CTL base oil and the VII displayed a lower CCS viscosity than that of mineral oil, suggesting it had better low-temperature performance and was able to quickly form a protective oil film on the surface, which was beneficial for the cold start. The functional group distribution state of the VII in base oil was analyzed using synchrotron radiation micro-infrared microscope (SR Micro-IR) technology, which revealed that HSD-1 had a better molecular interaction with CTL6 than 150N because of the better uniformity of the C=C group distribution. Based on this, a SP 0W-20 gasoline engine oil was developed by the combination of CTL base oil and the HSD-1 viscosity index improver, together with an additive package, a polymethacrylate pour point depressant, and a non-silicone defoamer, which showed excellent low-temperature performance, thermal oxidation stability, and detergency performance compared to the reference oil.

Nonlinear Current–Voltage Properties of Polymer Composite Doped With Calcium Copper Titanate Fibers

Since the discovery of gigantic permittivity properties of CaCu3Ti4O12 (CCTO) ceramics, development of its polymer composites application in supercapacitors, varistors and switching devices has been extensively investigated. Here the one-dimensional (1D) nano-sized CCTO nanofibers were prepared and used as fillers to improve the varistor Characteristics of epoxy composite materials. An obvious varistor effect was observed in CCTO_nfs/epoxy composite. The mechanisms of this varistor effect were analyzed by scanning capacitance microscopy technology. The nonlinear coefficient (α) of CCTO_nfs/epoxy composite with 48 wt% filler loading is about 4.65, which is much higher than of 2.42 for a single CCTO nanofiber. The mechanisms for this discrepancy were analyzed by scanning capacitance microscopy technology. The results show that the nonlinear current density-electric filed (J-E) relationship of the epoxy composites is contributed by the synergistic effects of potential barriers established at interfaces, grain boundaries and the participation of oxygen vacancies in CCTO nanofibers. All these results presented here provide a new insight into the electrical properties of CCTO. The CCTO_nfs/epoxy composites should have good application prospects in polymer composite varistor.

Understanding the unique corrosion phenomenon of Sanxingdui embrittled gold masks using scanning electrochemical cell microscopy (SECCM): Intergranular corrosion of Au-Ag-Pb ternary alloy

The study investigated the rare corrosion of embrittled gold masks excavated from the Sanxingdui archaeological site, attributing the embrittlement to preferential corrosion of intergranular Pb-containing phases through morphological studies using metallographic microscopy and SEM-EDS. Scanning electrochemical cell microscopy (SECCM) technology was used to quantitatively analyze simulated Au-Ag-Pb alloy samples to compare the electrochemical properties of different phases at the microscopic level. The results revealed an electrochemical activity order of Au-Ag < Au2Pb < AuPb2+PbxOy, which explains the varying levels of retention for different components in gold masks and provides a reference for subsequent conservation strategy.

Structures of Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus virions reveal species-specific tegument and envelope features.

Discovered in 1964, Epstein-Barr virus (EBV) is the first identified human oncogenic virus and the founding member of the gammaherpesvirus subfamily. In 1994, another cancer-causing virus was discovered in lesions of AIDS patients and later named Kaposi's sarcoma-associated herpesvirus (KSHV), the second human gammaherpesvirus. Despite the historical importance of EBV and KSHV, technical difficulties with isolating large quantities of these viruses and the pleiomorphic nature of their envelope and tegument layers have limited structural characterization of their virions. In this study, we employed the latest technologies in cryogenic electron microscopy (cryoEM) and tomography (cryoET) supplemented with an artificial intelligence-powered data processing software package to reconstruct 3D structures of the EBV and KSHV virions. We uncovered unique properties of the envelope glycoproteins and tegument layers of both EBV and KSHV. Comparison of these features with their non-tumorigenic counterparts provides insights into their relevance during infection.

Role of Emerin in regulating fibroblast differentiation and migration at the substrate of stiffness coupled topology.

In hypertrophic scars, the differentiation and migration of fibroblasts are influenced by the extracellular matrix microenvironment, which includes factors such as stiffness, restraint, and tensile force. These mechanical stresses incite alterations in cell behavior, accompanied by cytoskeletal protein reorganization. However, the role of nucleo-skeletal proteins in this context remains underexplored. In this study, we use a polyacrylamide hydrogel (PAA) to simulate the mechanical stress experienced by cells in scar tissue and investigate the impact of Emerin on cell behavior. We utilize atomic force microscopy (AFM) and RNA interference technology to analyze cell differentiation, migration, and stiffness. Our findings reveal that rigid substrates and cellular restriction elevate Emerin expression and diminish differentiation. Conversely, reducing Emerin expression leads to attenuated cell differentiation, where stiffness and constraining factors exert no notable influence. Furthermore, a softening of cells and an enhanced migration rate are also markedly observed. These observations indicate that variations in nuclear skeletal proteins, prompted by diverse matrix microenvironments, play a pivotal role in the pathogenesis of hypertrophic scars (HSs). This research offers novel insights and a reference point for understanding scar fibrosis formation mechanisms and preventing fibrosis.

Effect of grain structure on fatigue crack propagation behavior of 2024 aluminum alloy under different stress ratios

The fatigue load that a material experiences and its microstructure are important factors influencing fatigue crack propagation behavior. This study employed laser scanning microscopy and electron backscatter diffraction (EBSD) technology, along with fatigue crack propagation experiments, to investigate the fatigue crack propagation behavior of 2024 aluminum alloy under varying stress ratios (R). The results showed that the stress amplitude (σp) was the main factor controlling the fatigue crack propagation life (N). Additionally, detailed characterization of the fatigue crack propagation path was conducted using crystal models and EBSD. A fatigue crack propagation model for 2024 aluminum alloy under different R-values was established based on the grain twist angle and Schmid factor. Finally, the impact of R on the crack tip shielding (Ks) was systematically analyzed, elucidating the intrinsic mapping relationship between R, microstructure, and crack propagation characteristics.

A novel 3D instance segmentation network for synapse reconstruction from serial electron microscopy images

Synapses are fundamental components of how neurons communicate with each other and have attracted widespread attention from neuroscientists. Due to the rapid development of electron microscopy (EM) technology, imaging synapses at nanometer scale has become possible. However, the automation and efficacy of the synapse detection algorithm have not yet met expectations. The most popular approach involves a two-step process in which binary segmentation masks are first obtained and then connected components are used to produce reconstruction results. In this paper, an intelligent system to detect and segment synapses from serial section EM images is proposed. Specifically, a novel 3D instance segmentation network that can predict the synapses end-to-end is presented. The network can exploit and summarize features consistent with the biological structures of synapses, which is similar to the process of manual annotation. A block-wise inference strategy that adapts well to large-scale EM images is then introduced. Finally, two public datasets are used to evaluate our method. Experimental results demonstrate the superiority of the proposed approach, thus enabling computer-assisted analysis of synapses for neuroscientists.

Fast color fourier Ptychographic microscopic imaging technology with fusion color correction

In biomedicine, rapid and accurate acquisition of color pathology slides is of paramount importance for diagnostic analyses. Conventional microscopic full-color imaging cannot provide the physician with enough effective information at one time due to the "law of proportionality of objective lenses." Fourier ptychographic microscopy (FPM) has emerged as a solution for high-throughput digital pathology. It can simultaneously achieve a large field of view and high-resolution (HR) imaging. However, achieving color FPM typically requires capturing images separately for each of the red, green, and blue channels for reconstruction, resulting in a significant increase in the time cost. Existing improvement strategies require long coloring times, and have difficulty ensuring standardization between the reconstructed and original image colors owing to the influence of optical system stability and algorithm errors. In this study, a fast-color FPM imaging technology with fusion color correction (CCFPM) is proposed. It achieves high-precision color FPM imaging by separately capturing single low-resolution images for the red and blue channels, which are used to color the reconstructed green-channel HR grayscale image. The reconstructed HR color image is then processed using a color correction algorithm. Simulation and experimental analyses demonstrate that this method compresses the coloring algorithm's computation time to less than 1 s, reducing both the acquisition and reconstruction times by 66.7 % compared with the traditional R + G + B channel approach, achieving a significant improvement in speed. Meanwhile, it also achieves a breakthrough in algorithm speed compared with other fast color FPM solutions, compressing the coloring time to a few hundredths or even a few thousandths of the existing algorithms, and ensures the accuracy of the reconstructed image, reducing the RMSE by 50 % compared with the existing algorithms. Regarding color consistency, CCFPM can mitigate the chromatic aberration caused by fluctuations in optical system stability, maintaining consistency between the reconstructed and original image colors, thus achieving standardization of image color appearance. Therefore, the proposed method is promising for further advancements in the application of FPM in digital pathology.

A scalable and modular computational pipeline for axonal connectomics: automated tracing and assembly of axons across serial sections.

Progress in histological methods and in microscope technology has enabled dense staining and imaging of axons over large brain volumes, but tracing axons over such volumes requires new computational tools for 3D reconstruction of data acquired from serial sections. We have developed a computational pipeline for automated tracing and volume assembly of densely stained axons imaged over serial sections, which leverages machine learning-based segmentation to enable stitching and alignment with the axon traces themselves. We validated this segmentation-driven approach to volume assembly and alignment of individual axons over centimeter-scale serial sections and show the application of the output traces for analysis of local orientation and for proofreading over aligned volumes. The pipeline is scalable, and combined with recent advances in experimental approaches, should enable new studies of mesoscale connectivity and function over the whole human brain.

STIM1, ORAI1, and KDM2B in circulating tumor cells (CTCs) isolated from prostate cancer patients.

Introduction: Previous publications have shown that STIM1, ORAI1, and KDM2B, are implicated in Ca2+ signaling and are highly expressed in various cancer subtypes including prostate cancer. They play multiple roles in cancer cell migration, invasion, and metastasis. In the current study we investigated the expression of the above biomarkers in circulating tumor cells from patients with metastatic prostate cancer. Methods: Thirty-two patients were enrolled in this study and CTCs' isolation was performed with Ficoll density gradient. Two different triple immunofluorescence stainings were conducted with the following combination of antibodies: CK/KDM2B/CD45 and CK/STIM1/ORAI1. Slides were analyzed using VyCAP microscopy technology. Results: CTC-positive patients were detected in 41% for (CK/KDM2B/CD45) staining and in 56% for (CK/STIM1/ORAI1) staining. The (CK+/KDM2B+/CD45-) and the (CK+/STIM1+/ORAI1+) were the most frequent phenotypes as they were detected in 85% and 94% of the CTC-positive patients, respectively. Furthermore, the expression of ORAI1 and STIM1 in patients' PBMCs was very low exhibiting them as interesting specific biomarkers for CTC detection. The (CK+/STIM1+/ORAI1+) phenotype was correlated to bone metastasis (p = 0.034), while the (CK+/STIM1+/ORAI1-) to disease relapse (p = 0.049). Discussion: STIM1, ORAI1, and KDM2B were overexpressed in CTCs from patients with metastatic prostate cancer. STIM1 and ORAI1 expression was related to disease recurrence and bone metastasis. Further investigation of these biomarkers in a larger cohort of patients will clarify their clinical significance for prostate cancer patients.

Highly Efficient Fluorescent Probe to Visualize Protein Interactions at the Superresolution.

Superresolution microscopy (SR microscopy) of protein-protein interactions (PPIs) occurring in subcellular structures is essential for understanding cellular functions. However, a powerful and useful technology for SR microscopy of PPIs remains elusive. Here, we develop a highly efficient photoconvertible fluorescent probe, named split-Dendra2, for SR microscopy of PPIs in the cell. We found that split-Dendra2 enables a highly efficient detection of PPIs, making it possible to perform SR microscopy of PPIs with high spatial resolution and high image reconstruction fidelity. We demonstrate the utility of split-Dendra2 by visualizing PPIs occurring in small subcellular structures at the superresolution, such as clathrin-coated pits and focal adhesions, which cannot be visualized by the existing tools. Split-Dendra2 offers a powerful and useful tool that greatly expands the possibility of SR microscopy and can contribute to revealing the function of PPIs at the nanoscale resolution.

Fast, Accurate, and Versatile Data Analysis Platform for the Quantification of Molecular Spatiotemporal Signals.

Optical recording of intricate molecular dynamics is becoming an indispensable technique for biological studies, accelerated by the development of new or improved biosensors and microscopy technology. This creates major computational challenges to extract and quantify biologically meaningful spatiotemporal patterns embedded within complex and rich data sources, many of which cannot be captured with existing methods. Here, we introduce Activity Quantification and Analysis (AQuA2), a fast, accurate, and versatile data analysis platform built upon advanced machine learning techniques. It decomposes complex live imaging-based datasets into elementary signaling events, allowing accurate and unbiased quantification of molecular activities and identification of consensus functional units. We demonstrate applications across a wide range of biosensors, cell types, organs, animal models, and imaging modalities. As exemplar findings, we show how AQuA2 identified drug-dependent interactions between neurons and astroglia, and distinct sensorimotor signal propagation patterns in the mouse spinal cord.

Super-resolution Microscopy and Photo-lithography: How can one inspire the other

The principles of microscopy and lithography projection techniques are similar. To improve the resolution of the microscope or further reduce the image projection, both techniques face similar limitations, diffraction limits. The diffraction limit is a limitation of physical optics. In the development process of microscopes, to improve optical microscopy technology, researchers have proposed the idea of reducing the wavelength of light sources and increasing the numerical aperture based on the principle of diffraction limit generation. Modern scientists have proposed techniques to break through the diffraction limit and gradually increase the resolution of microscopes to further improve resolution. These breakthroughs also provide ideas for lithography technology, and researchers have invented techniques such as immersion lithography, multi-exposure, and two-photon direct writing. This article introduces the development process of super-resolution microscopy and lithography technology and analyzes the inspiration and influence of microscopy technology on lithography technology, that is, providing ideas for improving the accuracy of lithography technology.