Imaging-based phenotypic screening uses cellular phenotypes to describe the drug performance, which generally focuses on single cellular feature, lacking of a comprehensive characterization. Here, we propose a high co...Imaging-based phenotypic screening uses cellular phenotypes to describe the drug performance, which generally focuses on single cellular feature, lacking of a comprehensive characterization. Here, we propose a high content phenotypic screening method based on expansion microscopy(ExM) assisted cell painting, which enables multi-channel imaging with approximately 50 nm resolution. As a demonstration, we applied this method to a phenotypic screening involving five drugs. The morphological attributes of three subcellular structures were summarized to consist a “fingerprint” describing the drug effect. The proposed method can provide comprehensive and detailed clues for drug evaluation, enriching the content of phenotypic screening.展开更多
AIM:To investigate morphological changes in the cornea and meibomian glands using in vivo confocal microscopy(IVCM)in patients with type 2 diabetes(T2DM)and to analyze the correlation of these changes with disease dur...AIM:To investigate morphological changes in the cornea and meibomian glands using in vivo confocal microscopy(IVCM)in patients with type 2 diabetes(T2DM)and to analyze the correlation of these changes with disease duration(DD).METHODS:Patients with T2DM who visited the ophthalmology and endocrinology departments of our hospital from May 2023 to December 2023 were selected.According to DD,they were divided into the short-DD(≤10y)and long-DD(>10y)groups.The control group consisted of age-matched non-diabetes patients.All underwent Ocular Surface Disease Index(OSDI),tear break-up time(TBUT),Schirmer Ⅰ test(SIT),and IVCM imaging.Corneal nerve parameters,including corneal nerve fiber density(CNFD),corneal nerve branch density(CNBD),corneal nerve fiber length(CNFL),and meibomian gland metrics,including density of meibomian gland acinar units(MGAUD),the longest diameter of the meibomian gland acini(MGALD),the shortest diameter of meibomian gland acini(MGASD),the area of meibomian gland acinar units(MGAUA),were analyzed using ImageJ and ACCMetrics.RESULTS:A total of 130 patients with T2DM were included in this study,among which 68 were male(52.3%,age 30-76y)and 62 were female(47.7%,age 30-76y),the average age of the group was 55.54±11.65y.Significant differences(P0.05).CONCLUSION:T2DM patients exhibit greater corneal nerve and meibomian gland damage than age 30-76y nontype 2 diabetes patients.Prolonged DD exacerbates these morphological changes.展开更多
Microscopy imaging is fundamental in analyzing bacterial morphology and dynamics,offering critical insights into bacterial physiology and pathogenicity.Image segmentation techniques enable quantitative analysis of bac...Microscopy imaging is fundamental in analyzing bacterial morphology and dynamics,offering critical insights into bacterial physiology and pathogenicity.Image segmentation techniques enable quantitative analysis of bacterial structures,facilitating precise measurement of morphological variations and population behaviors at single-cell resolution.This paper reviews advancements in bacterial image segmentation,emphasizing the shift from traditional thresholding and watershed methods to deep learning-driven approaches.Convolutional neural networks(CNNs),U-Net architectures,and three-dimensional(3D)frameworks excel at segmenting dense biofilms and resolving antibiotic-induced morphological changes.These methods combine automated feature extraction with physics-informed postprocessing.Despite progress,challenges persist in computational efficiency,cross-species generalizability,and integration with multimodal experimental workflows.Future progress will depend on improving model robustness across species and imaging modalities,integrating multimodal data for phenotype-function mapping,and developing standard pipelines that link computational tools with clinical diagnostics.These innovations will expand microbial phenotyping beyond structural analysis,enabling deeper insights into bacterial physiology and ecological interactions.展开更多
Desulfurization of CaO–Al2O3 particles in molten steel was observed in situ using high-temperature confocal scanning laser microscopy.The effects of the aluminum and silicon contents of molten steel on desulfur...Desulfurization of CaO–Al2O3 particles in molten steel was observed in situ using high-temperature confocal scanning laser microscopy.The effects of the aluminum and silicon contents of molten steel on desulfurization were analyzed.When the total aluminum content in the steel increased from 6 to 1100 ppm,the CaS content in CaO–Al2O3 particles increased from 2.1wt%to 84.84wt%after the reaction for 90 s.Furthermore,when the silicon content in the steel increased from 0.01wt%to 2.20wt%,the CaS content in CaO–Al2O3 particles increased from 1.53wt%to 79.01wt%after the reaction for 90 s.This indicates that the increase in the aluminum and silicon contents of the steel promoted the desulfurization of CaO–Al2O3 particles.A kinetic model was established to predict the CaO–Al2O3 particles composition,and the diffusion coefficient of sulfur in CaO–Al2O3 particles was 9.375×10−10m2·s−1 at 1600℃,which provided a new method for the calculation of diffusion coefficient.展开更多
U-Net,a fully convolutional neural network(FCNN)with U-shaped features,has demonstrated significant success in biomedical image segmentation.However,the locality of convolution operations in the U-Net limits its abili...U-Net,a fully convolutional neural network(FCNN)with U-shaped features,has demonstrated significant success in biomedical image segmentation.However,the locality of convolution operations in the U-Net limits its ability to learn long-range dependencies.Transformers,originally developed for natural language processing,have recently been adapted for image segmentation because of their global self-attention mechanisms.Inspired by the long-range feature learning capability of transformers,we propose Dense-Transformer(DenT),an architecture designed for volumetric microscopy image segmentation.DenT incorporates transformers as encoders within each convolutional layer to capture global contextual information.Additionally,dense skip connections at multiple resolutions enhance feature propagation,enabling precise localization.We evaluated DenT on mitochondrial segmentation using our confocal microscopy dataset and a public fluorescence microscope dataset from the Allen Institute for Cell Science.The experimental results demonstrate that DenT incrementally improves the segmentation of mitochondria and mitochondrial DNA substructures from transmitted light microscopy images.DenT offers a tool for visualization,measurement,and analysis of mitochondrial morphology and mitochondrial DNA in label-free microscopy.展开更多
Structural anisotropy of celery stalks was studied using T2 anisotropy in microscopic MRI(μMRI)and supplemented by quantitative polarized light microscopy(PLM)at optical resolutions.Parenchyma,which is the ground ...Structural anisotropy of celery stalks was studied using T2 anisotropy in microscopic MRI(μMRI)and supplemented by quantitative polarized light microscopy(PLM)at optical resolutions.Parenchyma,which is the ground tissue in celery and has a diameter in the range of 50–90μm,was found to have isotropic T2;in contrast,collenchyma,which is the structural part of the ground tissue in celery and has a diameter in the range of 8–12μm,was found to have strong anisotropic T2.Substantial size variations within each porous structure and substantial co-existences of more than one type of structural tissues within a singleμMRI voxel were noticed in the optical images,which can contribute to the less clear anisotropies in the smaller vascular structures(e.g.,phloem(approximately 2–6μm in diameter)and xylem(approximately 5–15μm in diameter)).Celery could be used as a simple plant model to study the relationships between tissue microstructures and nuclear spin relaxation in fibrous and porous specimens.展开更多
By 2024,Malaysia achieved a significant public health milestone by marking its seventh consecutive year without a single local transmission of indigenous human Plasmodium species.The country has sustained elimination ...By 2024,Malaysia achieved a significant public health milestone by marking its seventh consecutive year without a single local transmission of indigenous human Plasmodium species.The country has sustained elimination of non-zoonotic malaria,primarily caused by Plasmodium(P.)falciparum and P.vivax,while remaining vigilant against imported and introduced cases driven by cross-border movement and international travel.However,despite this achievement,Malaysia has emerged as the global epicentre of zoonotic for P.knowlesi.展开更多
In recent years,magic-angle twisted bilayer graphene has emerged as a focal point in condensed matter physics due to its novel quantum properties arising from ultra-flat electronic bands.Subsequent research has also r...In recent years,magic-angle twisted bilayer graphene has emerged as a focal point in condensed matter physics due to its novel quantum properties arising from ultra-flat electronic bands.Subsequent research has also revealed analogous flat-band physics in other twisted multilayer graphene systems,such as twisted monolayer-bilayer graphene,twisted trilayer graphene,and twisted double-bilayer graphene.These graphene-based twisted systems host a rich variety of strongly correlated and topological quantum states—including ferromagnetism,superconductivity,Chern insulators,and the quantum anomalous Hall effect—significantly advancing the investigation of strongly correlated and topological physics in real materials.Here,we conduct a systematic review of the recent advances in the exploration of flat-band and strong correlation physics using scanning tunneling microscopy/spectroscopy in magic-angle twisted multilayer graphene.First,we briefly introduce the theoretical background of the flat-band physics in magic-angle twisted multilayer graphene.Subsequently,we present in detail the strongly correlated electronic states and exotic quantum phenomena observed by scanning tunneling microscopy/spectroscopy in various twisted graphene platforms,including twisted bilayer graphene,twisted monolayer−bilayer graphene,twisted trilayer graphene,and twisted double-bilayer graphene.Lastly,we provide a summary and an outlook for future research in this field.展开更多
Light-field fluorescence microscopy(LFM)enables high-speed volumetric imaging by capturing 3D information in a single snapshot.However,its reconstruction quality is severely degraded by detection noise.We propose SSD-...Light-field fluorescence microscopy(LFM)enables high-speed volumetric imaging by capturing 3D information in a single snapshot.However,its reconstruction quality is severely degraded by detection noise.We propose SSD-LFM,a self-supervised denoising framework that incorporates physical models,to substantially remove noise-induced artifacts in multimodality LFM.The SSD-LFM model is trained exclusively on low-signal-to-noise ratio(SNR)data,bypassing the need for paired noisy-clean datasets for network training.The denoising performance of SSD-LFM is validated on conventional LFM,scanning LFM(sLFM),and two-photon excitation LFM(TP-LFM).Comprehensive evaluations on both simulated and experimental data demonstrate that SSD-LFM achieves artifact-free 3D reconstruction with over 5-fold fewer photons.For time-lapse imaging,the self-supervised architecture further enables SSD-LFM to operate only on the collected noisy raw data,overcoming generalization constraints inherent in supervised methods.Due to the significantly improved quality under low-SNR excitation,SSD-LFM enables long-term volumetric observation of phototoxicity-vulnerable bioprocesses.展开更多
Ever since its inception,axially swept light-sheet microscopy(ASLM)has demonstrated its diverse imaging capability,particularly in the context of large,cleared specimens.Yet current implementations face fundamental li...Ever since its inception,axially swept light-sheet microscopy(ASLM)has demonstrated its diverse imaging capability,particularly in the context of large,cleared specimens.Yet current implementations face fundamental limitations in optimizing imaging performance for heterogeneous samples.Although significant advances have been made in ASLM development,obtaining sufficient signal while maintaining optimal axial resolution remains a persistent challenge.Moreover,current ASLM implementations use fixed illumination numerical apertures(NAs)or lossy mechanical slits,preventing dynamic light-sheet waist control,thereby restricting multiscale imaging flexibility.Here,we present an ASLM-based optical method that enhances sample signal detection without the need to increase laser power while providing flexibility in axial resolution control ranging from 500 nm to 1.6μm.Our approach enables flexible and lossless control of light-sheet thickness in the sample space to achieve variable axial resolution within the specified range.This tunable beam shaping capability addresses the fundamental limitations of current ASLM systems by providing researchers with the ability to optimize imaging parameters by bridging the critical gap between resolution and signal in ASLM-based microscopy.We demonstrate the utility of our approach by imaging fluorescent beads and various cleared-tissue samples.展开更多
Compressed streak imaging(CSI)is a computational imaging strategy that can acquire video at over 150 trillion frames per second.Despite this achievement,CSI faces challenges in detecting subtle intensity fluctuations ...Compressed streak imaging(CSI)is a computational imaging strategy that can acquire video at over 150 trillion frames per second.Despite this achievement,CSI faces challenges in detecting subtle intensity fluctuations in slow-moving,continuously illuminated objects.This limitation,largely attributable to high streak compression and motion blur,has curtailed the broader adoption of CSI in cellular fluorescence microscopy.To address these issues and expand the utility of CSI,we developed a two-axis CSI(TACSI)method that results in significant improvements to the reconstructed video fidelity.TACSI introduces a second scanning axis,which shuttles a conjugate image of the object with respect to the coded aperture(CA).The moving image decreases the localized spatiotemporal mixing and produces a“flash and shutter”phenomenon that reduces CA motion blur,overcoming the limitations of current CSI technologies.This approach is supported by an analytical model describing the TACSI compression ratio,along with simulated and empirical measurements.We demonstrate TACSI’s ability tomeasure rapid variations in cellmembrane potentials,previously unattainable with conventional CSI.This method has broad implications for high-speed photography,including visualization of action potentials,muscle contractions,and enzymatic reactions that occur on microsecond and faster timescales using fluorescence microscopy.展开更多
The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are revi...The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are reviewed and summarized,particularly the col-lision of various inclusions,dissolution of inclusions in liquid slag,and reactions between inclusions and steel.Solid inclusions exhibited a high collision tendency,whereas pure liquid inclusions exhibited minimal collisions because of the small attraction force induced by their<90°contact angle with molten steel.The collision of complex inclusions in molten steel was not included in the scope of this study and should be evaluated in future studies.Higher CaO/Al2O3and CaO/SiO2ratios in liquid slag promoted the dissolution of Al2O3-based in-clusions.The formation of solid phases in the slag should be prevented to improve dissolution of inclusions.To accurately simulate the dissolution of inclusions in liquid slag,in-situ observation of the dissolution of inclusions at the steel-slag interface is necessary.Using a combination of CSLM and scanning electron microscopy-energy dispersive spectroscopy,the composition and morphological evolution of the inclusions during their modification by the dissolved elements in steel were observed and analyzed.Although the in-situ observa-tion of MnS and TiN precipitations has been widely studied,the in-situ observation of the evolution of oxide inclusions in steel during so-lidification and heating processes has rarely been reported.The effects of temperature,heating and cooling rates,and inclusion character-istics on the formation of acicular ferrites(AFs)have been widely studied.At a cooling rate of 3-5 K/s,the order of AF growth rate in-duced by different inclusions,as reported in literature,is Ti-O<Ti-Ca-Zr-Al-O<Mg-O<Ti-Zr-Al-O<Mn-Ti-Al-O<Ti-Al-O<Zr-Ti-Al-O.Further comprehensive experiments are required to investigate the quantitative relationship between the formation of AFs and inclusions.展开更多
Fourier ptychographic microscopy(FPM)is an innovative computational microscopy approach that enables high-throughput imaging with high resolution,wide field of view,and quantitative phase imaging(QPI)by simultaneously...Fourier ptychographic microscopy(FPM)is an innovative computational microscopy approach that enables high-throughput imaging with high resolution,wide field of view,and quantitative phase imaging(QPI)by simultaneously capturing bright-field and dark-field images.However,effectively utilizing dark-field intensity images,including both normally exposed and overexposed data,which contain valuable high-angle illumination information,remains a complex challenge.Successfully extracting and applying this information could significantly enhance phase reconstruction,benefiting processes such as virtual staining and QPI imaging.To address this,we introduce a multi-exposure image fusion(MEIF)framework that optimizes dark-field information by incorporating it into the FPM preprocessing workflow.MEIF increases the data available for reconstruction without requiring changes to the optical setup.We evaluate the framework using both feature-domain and traditional FPM,demonstrating that it achieves substantial improvements in intensity resolution and phase information for biological samples that exceed the performance of conventional high dynamic range(HDR)methods.This image preprocessing-based information-maximization strategy fully leverages existing datasets and offers promising potential to drive advancements in fields such as microscopy,remote sensing,and crystallography.展开更多
To investigate the mechanisms underlying the onset and progression of ischemic stroke,some methods have been proposed that can simultaneously monitor and create embolisms in the animal cerebral cortex.However,these me...To investigate the mechanisms underlying the onset and progression of ischemic stroke,some methods have been proposed that can simultaneously monitor and create embolisms in the animal cerebral cortex.However,these methods often require complex systems and the effect of age on cerebral embolism has not been adequately studied,although ischemic stroke is strongly age-related.In this study,we propose an optical-resolution photoacoustic microscopy-based visualized photothrombosis methodology to create and monitor ischemic stroke in mice simultaneously using a 532 nm pulsed laser.We observed the molding process in mice of different ages and presented age-dependent vascular embolism differentiation.Moreover,we integrated optical coherence tomography angiography to investigate age-associated trends in cerebrovascular variability following a stroke.Our imaging data and quantitative analyses underscore the differential cerebrovascular responses to stroke in mice of different ages,thereby highlighting the technique's potential for evaluating cerebrovascular health and unraveling age-related mechanisms involved in ischemic strokes.展开更多
The infinite-layer nickelates,proposed as analogs to superconducting cuprates,provide a promising platform for exploring the mechanisms of unconventional superconductivity.However,the superconductivity has been exclus...The infinite-layer nickelates,proposed as analogs to superconducting cuprates,provide a promising platform for exploring the mechanisms of unconventional superconductivity.However,the superconductivity has been exclusively observed in thin films under atmospheric pressure,underscoring the critical role of the heterointerface.展开更多
Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often ...Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often posing diagnostic challenges.Dermoscopy and reflectance confocal microscopy(RCM)can aid in their differentiation,yet their specific features under these tools have not been systematically described.This study aims to summarize and analyze the dermoscopic and RCM features of VEN,SK,VP,VV,and NS.Methods:A total of 121 patients with histopathologically confirmed verrucous proliferative skin diseases were enrolled.Dermoscopy and RCM imaging was used to observe and analyze the microscopic features of these conditions.Results:Under dermoscopy,the 5 diseases displayed distinct characteristics:VEN typically showed gyriform structures;SK was characterized by gyriform structures,comedo-like openings,and milia-like cysts;VP and VV featured dotted vessels and frogspawn-like structures;NS presented as brownish-yellow globules.RCM revealed shared features such as hyperkeratosis and acanthosis across all 5 diseases.Specific features included gyriform structures and elongated rete ridges in VEN;pseudocysts and gyriform structures in SK;evenly distributed ring-like structures in VP;vacuolated cells and papillomatous proliferation in VV;and frogspawn-like structures in NS.Conclusion:These 5 verrucous proliferative skin conditions exhibit distinguishable features under both dermoscopy and RCM.The combination of these 2 noninvasive imaging modalities holds significant clinical value for the differential diagnosis of verrucous proliferative skin diseases.展开更多
Digital in-line holographic microscopy(DIHM)is a non-invasive,real-time,label-free technique that captures three-dimensional(3D)positional,orientational,and morphological information from digital holographic images of...Digital in-line holographic microscopy(DIHM)is a non-invasive,real-time,label-free technique that captures three-dimensional(3D)positional,orientational,and morphological information from digital holographic images of living biological cells.Unlike conventional microscopies,the DIHM technique enables precise measurements of dynamic behaviors exhibited by living cells within a 3D volume.This review outlines the fundamental principles and comprehensive digital image processing procedures employed in DIHM-based cell tracking methods.In addition,recent applications of DIHM technique for label-free identification and digital tracking of various motile biological cells,including human blood cells,spermatozoa,diseased cells,and unicellular microorganisms,are thoroughly examined.Leveraging artificial intelligence has significantly enhanced both the speed and accuracy of digital image processing for cell tracking and identification.The quantitative data on cell morphology and dynamics captured by DIHM can effectively elucidate the underlying mechanisms governing various microbial behaviors and contribute to the accumulation of diagnostic databases and the development of clinical treatments.展开更多
Recently,charged solitons have been found in a two-dimensional CoCl2/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl2 using scanning ...Recently,charged solitons have been found in a two-dimensional CoCl2/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl2 using scanning tunneling microscopy(STM)and atomic force microscopy(AFM).Moreover,we study the electrical properties of the charged solitons at zero electric field by measuring local contact potential difference(LCPD)via Kelvin probe force microscopy(KPFM)using the Δf(V)method.The compensation voltage corresponding to the vertex of the parabola is obtained by fitting the quadratic relationship between Δf and sample bias.The results show that,without an external electric field,the solitons behave as negatively charged entities.Meanwhile,the LCPD mapping characterizes the spatial distribution of the potential at the charged solitons,which agrees well with those obtained from STM band bending measurements.展开更多
The photocatalytic activity of a photocatalyst is significantly influenced by its microstructure.Therefore,it is crucial to understand and characterize the microstructure.Transmission electron microscopy(TEM)has been ...The photocatalytic activity of a photocatalyst is significantly influenced by its microstructure.Therefore,it is crucial to understand and characterize the microstructure.Transmission electron microscopy(TEM)has been an indispensable tool that provides direct visualization of the microstructure at the atomic level.Despite numerous reviews on photocatalytic processes,there has been a lack of comprehensive summaries that focus on the critical role of TEM in photocatalytic applications.This review aims to fill this gap by highlighting the significant contributions of TEM techniques in understanding key photocatalytic processes,including light absorption,charge separation and transfer,and surface reaction.In addition,this review discusses in-situ TEM techniques for observing photocatalytic reactions and explores the challenges and future perspectives related to the application of TEM in photocatalysis.展开更多
基金supported by the National Key R&D Program of China(No.2022YFC2409603)the Tianjin Municipal Science and Technology Program(No.23JCZXJC00340)the Beijing-Tianjin-Hebei Fundamental Research Cooperation Project(No.J230001)。
摘要Imaging-based phenotypic screening uses cellular phenotypes to describe the drug performance, which generally focuses on single cellular feature, lacking of a comprehensive characterization. Here, we propose a high content phenotypic screening method based on expansion microscopy(ExM) assisted cell painting, which enables multi-channel imaging with approximately 50 nm resolution. As a demonstration, we applied this method to a phenotypic screening involving five drugs. The morphological attributes of three subcellular structures were summarized to consist a “fingerprint” describing the drug effect. The proposed method can provide comprehensive and detailed clues for drug evaluation, enriching the content of phenotypic screening.
基金Supported by 2019 Medical Science Research Key Project Plan of Hebei Province(No.201900341)2024 Medical Science Research Key Project Plan of Hebei Province(No.20242157)+2 种基金the Natual Science Foundation of Hebei Province(No.H2020206650)Hebei Provincial Government Funded Clinical Medicine Excellent Talents ProjectHebei Province Medical Application Technology Tracking Project(No.GZ2023091).
摘要AIM:To investigate morphological changes in the cornea and meibomian glands using in vivo confocal microscopy(IVCM)in patients with type 2 diabetes(T2DM)and to analyze the correlation of these changes with disease duration(DD).METHODS:Patients with T2DM who visited the ophthalmology and endocrinology departments of our hospital from May 2023 to December 2023 were selected.According to DD,they were divided into the short-DD(≤10y)and long-DD(>10y)groups.The control group consisted of age-matched non-diabetes patients.All underwent Ocular Surface Disease Index(OSDI),tear break-up time(TBUT),Schirmer Ⅰ test(SIT),and IVCM imaging.Corneal nerve parameters,including corneal nerve fiber density(CNFD),corneal nerve branch density(CNBD),corneal nerve fiber length(CNFL),and meibomian gland metrics,including density of meibomian gland acinar units(MGAUD),the longest diameter of the meibomian gland acini(MGALD),the shortest diameter of meibomian gland acini(MGASD),the area of meibomian gland acinar units(MGAUA),were analyzed using ImageJ and ACCMetrics.RESULTS:A total of 130 patients with T2DM were included in this study,among which 68 were male(52.3%,age 30-76y)and 62 were female(47.7%,age 30-76y),the average age of the group was 55.54±11.65y.Significant differences(P0.05).CONCLUSION:T2DM patients exhibit greater corneal nerve and meibomian gland damage than age 30-76y nontype 2 diabetes patients.Prolonged DD exacerbates these morphological changes.
基金financially supported by the Open Project Program of Wuhan National Laboratory for Optoelectronics(No.2022WNLOKF009)the National Natural Science Foundation of China(No.62475216)+2 种基金the Key Research and Development Program of Shaanxi(No.2024GH-ZDXM-37)the Fujian Provincial Natural Science Foundation of China(No.2024J01060)the Startup Program of XMU,and the Fundamental Research Funds for the Central Universities.
摘要Microscopy imaging is fundamental in analyzing bacterial morphology and dynamics,offering critical insights into bacterial physiology and pathogenicity.Image segmentation techniques enable quantitative analysis of bacterial structures,facilitating precise measurement of morphological variations and population behaviors at single-cell resolution.This paper reviews advancements in bacterial image segmentation,emphasizing the shift from traditional thresholding and watershed methods to deep learning-driven approaches.Convolutional neural networks(CNNs),U-Net architectures,and three-dimensional(3D)frameworks excel at segmenting dense biofilms and resolving antibiotic-induced morphological changes.These methods combine automated feature extraction with physics-informed postprocessing.Despite progress,challenges persist in computational efficiency,cross-species generalizability,and integration with multimodal experimental workflows.Future progress will depend on improving model robustness across species and imaging modalities,integrating multimodal data for phenotype-function mapping,and developing standard pipelines that link computational tools with clinical diagnostics.These innovations will expand microbial phenotyping beyond structural analysis,enabling deeper insights into bacterial physiology and ecological interactions.
基金supported by the National Key R&D Program of China(No.2023YFB3709900)the National Nature Science Foundation of China(No.U22A20171)+1 种基金the China Baowu Low Carbon Metallurgy Innovation Foundation(No.BWLCF202315)the High Steel Center(HSC)at North China University of Technology and University of Science and Technology Beijing,China.
摘要Desulfurization of CaO–Al2O3 particles in molten steel was observed in situ using high-temperature confocal scanning laser microscopy.The effects of the aluminum and silicon contents of molten steel on desulfurization were analyzed.When the total aluminum content in the steel increased from 6 to 1100 ppm,the CaS content in CaO–Al2O3 particles increased from 2.1wt%to 84.84wt%after the reaction for 90 s.Furthermore,when the silicon content in the steel increased from 0.01wt%to 2.20wt%,the CaS content in CaO–Al2O3 particles increased from 1.53wt%to 79.01wt%after the reaction for 90 s.This indicates that the increase in the aluminum and silicon contents of the steel promoted the desulfurization of CaO–Al2O3 particles.A kinetic model was established to predict the CaO–Al2O3 particles composition,and the diffusion coefficient of sulfur in CaO–Al2O3 particles was 9.375×10−10m2·s−1 at 1600℃,which provided a new method for the calculation of diffusion coefficient.
基金Taiwan University Center for Advanced Computing and Imaging in Biomedicine(NTU-114L900701).
摘要U-Net,a fully convolutional neural network(FCNN)with U-shaped features,has demonstrated significant success in biomedical image segmentation.However,the locality of convolution operations in the U-Net limits its ability to learn long-range dependencies.Transformers,originally developed for natural language processing,have recently been adapted for image segmentation because of their global self-attention mechanisms.Inspired by the long-range feature learning capability of transformers,we propose Dense-Transformer(DenT),an architecture designed for volumetric microscopy image segmentation.DenT incorporates transformers as encoders within each convolutional layer to capture global contextual information.Additionally,dense skip connections at multiple resolutions enhance feature propagation,enabling precise localization.We evaluated DenT on mitochondrial segmentation using our confocal microscopy dataset and a public fluorescence microscope dataset from the Allen Institute for Cell Science.The experimental results demonstrate that DenT incrementally improves the segmentation of mitochondria and mitochondrial DNA substructures from transmitted light microscopy images.DenT offers a tool for visualization,measurement,and analysis of mitochondrial morphology and mitochondrial DNA in label-free microscopy.
基金supported by a BSF grant from the United States-Israel Binational Science Foundation(2019033)a grant from the National Institutes of Health(R01 AR 69047).
摘要Structural anisotropy of celery stalks was studied using T2 anisotropy in microscopic MRI(μMRI)and supplemented by quantitative polarized light microscopy(PLM)at optical resolutions.Parenchyma,which is the ground tissue in celery and has a diameter in the range of 50–90μm,was found to have isotropic T2;in contrast,collenchyma,which is the structural part of the ground tissue in celery and has a diameter in the range of 8–12μm,was found to have strong anisotropic T2.Substantial size variations within each porous structure and substantial co-existences of more than one type of structural tissues within a singleμMRI voxel were noticed in the optical images,which can contribute to the less clear anisotropies in the smaller vascular structures(e.g.,phloem(approximately 2–6μm in diameter)and xylem(approximately 5–15μm in diameter)).Celery could be used as a simple plant model to study the relationships between tissue microstructures and nuclear spin relaxation in fibrous and porous specimens.
基金supported by the Geran Fundamental Fakulti Perubatan(FF-2025-202),Universiti Kebangsaan Malaysia.
摘要By 2024,Malaysia achieved a significant public health milestone by marking its seventh consecutive year without a single local transmission of indigenous human Plasmodium species.The country has sustained elimination of non-zoonotic malaria,primarily caused by Plasmodium(P.)falciparum and P.vivax,while remaining vigilant against imported and introduced cases driven by cross-border movement and international travel.However,despite this achievement,Malaysia has emerged as the global epicentre of zoonotic for P.knowlesi.
基金supported by the National Natural Science Foundation of China(Grant Nos.12474166 and 12174095)the Natural Science Foundation of Hunan Province,China(Grant No.2025JJ20001).
摘要In recent years,magic-angle twisted bilayer graphene has emerged as a focal point in condensed matter physics due to its novel quantum properties arising from ultra-flat electronic bands.Subsequent research has also revealed analogous flat-band physics in other twisted multilayer graphene systems,such as twisted monolayer-bilayer graphene,twisted trilayer graphene,and twisted double-bilayer graphene.These graphene-based twisted systems host a rich variety of strongly correlated and topological quantum states—including ferromagnetism,superconductivity,Chern insulators,and the quantum anomalous Hall effect—significantly advancing the investigation of strongly correlated and topological physics in real materials.Here,we conduct a systematic review of the recent advances in the exploration of flat-band and strong correlation physics using scanning tunneling microscopy/spectroscopy in magic-angle twisted multilayer graphene.First,we briefly introduce the theoretical background of the flat-band physics in magic-angle twisted multilayer graphene.Subsequently,we present in detail the strongly correlated electronic states and exotic quantum phenomena observed by scanning tunneling microscopy/spectroscopy in various twisted graphene platforms,including twisted bilayer graphene,twisted monolayer−bilayer graphene,twisted trilayer graphene,and twisted double-bilayer graphene.Lastly,we provide a summary and an outlook for future research in this field.
基金supported by the Natural Science Foundation of Beijing Municipality(Grant No.4254070)the National Natural Science Foundation of China(Grant No.62501030).
摘要Light-field fluorescence microscopy(LFM)enables high-speed volumetric imaging by capturing 3D information in a single snapshot.However,its reconstruction quality is severely degraded by detection noise.We propose SSD-LFM,a self-supervised denoising framework that incorporates physical models,to substantially remove noise-induced artifacts in multimodality LFM.The SSD-LFM model is trained exclusively on low-signal-to-noise ratio(SNR)data,bypassing the need for paired noisy-clean datasets for network training.The denoising performance of SSD-LFM is validated on conventional LFM,scanning LFM(sLFM),and two-photon excitation LFM(TP-LFM).Comprehensive evaluations on both simulated and experimental data demonstrate that SSD-LFM achieves artifact-free 3D reconstruction with over 5-fold fewer photons.For time-lapse imaging,the self-supervised architecture further enables SSD-LFM to operate only on the collected noisy raw data,overcoming generalization constraints inherent in supervised methods.Due to the significantly improved quality under low-SNR excitation,SSD-LFM enables long-term volumetric observation of phototoxicity-vulnerable bioprocesses.
基金supported by the National Institutes of Health(Grant Nos.R35GM151152 to T.C.,2P20GM130422-06 to O.P.,and R01CA293942 to R.E.S.)support was provided by the NSF career award(Grant No.2440821)to T.C.
摘要Ever since its inception,axially swept light-sheet microscopy(ASLM)has demonstrated its diverse imaging capability,particularly in the context of large,cleared specimens.Yet current implementations face fundamental limitations in optimizing imaging performance for heterogeneous samples.Although significant advances have been made in ASLM development,obtaining sufficient signal while maintaining optimal axial resolution remains a persistent challenge.Moreover,current ASLM implementations use fixed illumination numerical apertures(NAs)or lossy mechanical slits,preventing dynamic light-sheet waist control,thereby restricting multiscale imaging flexibility.Here,we present an ASLM-based optical method that enhances sample signal detection without the need to increase laser power while providing flexibility in axial resolution control ranging from 500 nm to 1.6μm.Our approach enables flexible and lossless control of light-sheet thickness in the sample space to achieve variable axial resolution within the specified range.This tunable beam shaping capability addresses the fundamental limitations of current ASLM systems by providing researchers with the ability to optimize imaging parameters by bridging the critical gap between resolution and signal in ASLM-based microscopy.We demonstrate the utility of our approach by imaging fluorescent beads and various cleared-tissue samples.
基金Work contributed by SAIC was performed under United States Air Force Contract No.FA8650-19-C-6024funding from the Air Force Office of Scientific Research under Award No.23RHCOR002+3 种基金funding from the Natural Sciences and Engineering Research Council of Canada(Grant No.RGPIN-2024-05551)funding from the Air Force Office of Scientific Research(Grant Nos.FA9550-20-1-0366,FA9550-20-1-0367,and FA9550-23-1-0599)the National Institutes of Health(NIH)(Grant Nos.R01GM127696,R01GM152633,R21GM142107,and 1R21CA269099)was supported by NASA,BARDA,NIH,and USFDA,under Contract/Agreement No.80ARC023CA002.
摘要Compressed streak imaging(CSI)is a computational imaging strategy that can acquire video at over 150 trillion frames per second.Despite this achievement,CSI faces challenges in detecting subtle intensity fluctuations in slow-moving,continuously illuminated objects.This limitation,largely attributable to high streak compression and motion blur,has curtailed the broader adoption of CSI in cellular fluorescence microscopy.To address these issues and expand the utility of CSI,we developed a two-axis CSI(TACSI)method that results in significant improvements to the reconstructed video fidelity.TACSI introduces a second scanning axis,which shuttles a conjugate image of the object with respect to the coded aperture(CA).The moving image decreases the localized spatiotemporal mixing and produces a“flash and shutter”phenomenon that reduces CA motion blur,overcoming the limitations of current CSI technologies.This approach is supported by an analytical model describing the TACSI compression ratio,along with simulated and empirical measurements.We demonstrate TACSI’s ability tomeasure rapid variations in cellmembrane potentials,previously unattainable with conventional CSI.This method has broad implications for high-speed photography,including visualization of action potentials,muscle contractions,and enzymatic reactions that occur on microsecond and faster timescales using fluorescence microscopy.
基金supported by the National Key R&D Program(No.2023YFB3709900)the National Nature Science Foundation of China(No.U22A20171)+2 种基金China Baowu Low Carbon Metallurgy Innovation Foundation(No.BWLCF202315)the High Steel Center(HSC)at North China University of TechnologyUniversity of Science and Technology Beijing,China.
摘要The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are reviewed and summarized,particularly the col-lision of various inclusions,dissolution of inclusions in liquid slag,and reactions between inclusions and steel.Solid inclusions exhibited a high collision tendency,whereas pure liquid inclusions exhibited minimal collisions because of the small attraction force induced by their<90°contact angle with molten steel.The collision of complex inclusions in molten steel was not included in the scope of this study and should be evaluated in future studies.Higher CaO/Al2O3and CaO/SiO2ratios in liquid slag promoted the dissolution of Al2O3-based in-clusions.The formation of solid phases in the slag should be prevented to improve dissolution of inclusions.To accurately simulate the dissolution of inclusions in liquid slag,in-situ observation of the dissolution of inclusions at the steel-slag interface is necessary.Using a combination of CSLM and scanning electron microscopy-energy dispersive spectroscopy,the composition and morphological evolution of the inclusions during their modification by the dissolved elements in steel were observed and analyzed.Although the in-situ observa-tion of MnS and TiN precipitations has been widely studied,the in-situ observation of the evolution of oxide inclusions in steel during so-lidification and heating processes has rarely been reported.The effects of temperature,heating and cooling rates,and inclusion character-istics on the formation of acicular ferrites(AFs)have been widely studied.At a cooling rate of 3-5 K/s,the order of AF growth rate in-duced by different inclusions,as reported in literature,is Ti-O<Ti-Ca-Zr-Al-O<Mg-O<Ti-Zr-Al-O<Mn-Ti-Al-O<Ti-Al-O<Zr-Ti-Al-O.Further comprehensive experiments are required to investigate the quantitative relationship between the formation of AFs and inclusions.
基金supported by the National Natural Science Foundation of China(Grant No.12104500)the Key Research and Development Projects of Shaanxi Province of China(Grant No.2023-YBSF-263).
摘要Fourier ptychographic microscopy(FPM)is an innovative computational microscopy approach that enables high-throughput imaging with high resolution,wide field of view,and quantitative phase imaging(QPI)by simultaneously capturing bright-field and dark-field images.However,effectively utilizing dark-field intensity images,including both normally exposed and overexposed data,which contain valuable high-angle illumination information,remains a complex challenge.Successfully extracting and applying this information could significantly enhance phase reconstruction,benefiting processes such as virtual staining and QPI imaging.To address this,we introduce a multi-exposure image fusion(MEIF)framework that optimizes dark-field information by incorporating it into the FPM preprocessing workflow.MEIF increases the data available for reconstruction without requiring changes to the optical setup.We evaluate the framework using both feature-domain and traditional FPM,demonstrating that it achieves substantial improvements in intensity resolution and phase information for biological samples that exceed the performance of conventional high dynamic range(HDR)methods.This image preprocessing-based information-maximization strategy fully leverages existing datasets and offers promising potential to drive advancements in fields such as microscopy,remote sensing,and crystallography.
基金supported by University of Macao,China,Nos.MYRG2022-00054-FHS and MYRG-GRG2023-00038-FHS-UMDF(to ZY)the Macao Science and Technology Development Fund,China,Nos.FDCT0048/2021/AGJ and FDCT0020/2019/AMJ and FDCT 0011/2018/A1(to ZY)Natural Science Foundation of Guangdong Province of China,No.EF017/FHS-YZ/2021/GDSTC(to ZY)。
摘要To investigate the mechanisms underlying the onset and progression of ischemic stroke,some methods have been proposed that can simultaneously monitor and create embolisms in the animal cerebral cortex.However,these methods often require complex systems and the effect of age on cerebral embolism has not been adequately studied,although ischemic stroke is strongly age-related.In this study,we propose an optical-resolution photoacoustic microscopy-based visualized photothrombosis methodology to create and monitor ischemic stroke in mice simultaneously using a 532 nm pulsed laser.We observed the molding process in mice of different ages and presented age-dependent vascular embolism differentiation.Moreover,we integrated optical coherence tomography angiography to investigate age-associated trends in cerebrovascular variability following a stroke.Our imaging data and quantitative analyses underscore the differential cerebrovascular responses to stroke in mice of different ages,thereby highlighting the technique's potential for evaluating cerebrovascular health and unraveling age-related mechanisms involved in ischemic strokes.
基金supported by the National Natural Science Foundation of China[52125307(to P.G.),12404192(to R.C.S),12274061(to L.Q.)]Key Research and Development Program from the Ministry of Science and Technology(2023YFA1406301)the support from the New Cornerstone Science Foundation through the XPLORER PRIZE。
摘要The infinite-layer nickelates,proposed as analogs to superconducting cuprates,provide a promising platform for exploring the mechanisms of unconventional superconductivity.However,the superconductivity has been exclusively observed in thin films under atmospheric pressure,underscoring the critical role of the heterointerface.
基金supported by the Project of Health Committee of Hunan Province(D202304128868),China.
摘要Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often posing diagnostic challenges.Dermoscopy and reflectance confocal microscopy(RCM)can aid in their differentiation,yet their specific features under these tools have not been systematically described.This study aims to summarize and analyze the dermoscopic and RCM features of VEN,SK,VP,VV,and NS.Methods:A total of 121 patients with histopathologically confirmed verrucous proliferative skin diseases were enrolled.Dermoscopy and RCM imaging was used to observe and analyze the microscopic features of these conditions.Results:Under dermoscopy,the 5 diseases displayed distinct characteristics:VEN typically showed gyriform structures;SK was characterized by gyriform structures,comedo-like openings,and milia-like cysts;VP and VV featured dotted vessels and frogspawn-like structures;NS presented as brownish-yellow globules.RCM revealed shared features such as hyperkeratosis and acanthosis across all 5 diseases.Specific features included gyriform structures and elongated rete ridges in VEN;pseudocysts and gyriform structures in SK;evenly distributed ring-like structures in VP;vacuolated cells and papillomatous proliferation in VV;and frogspawn-like structures in NS.Conclusion:These 5 verrucous proliferative skin conditions exhibit distinguishable features under both dermoscopy and RCM.The combination of these 2 noninvasive imaging modalities holds significant clinical value for the differential diagnosis of verrucous proliferative skin diseases.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT,RS-2023-00218630).
摘要Digital in-line holographic microscopy(DIHM)is a non-invasive,real-time,label-free technique that captures three-dimensional(3D)positional,orientational,and morphological information from digital holographic images of living biological cells.Unlike conventional microscopies,the DIHM technique enables precise measurements of dynamic behaviors exhibited by living cells within a 3D volume.This review outlines the fundamental principles and comprehensive digital image processing procedures employed in DIHM-based cell tracking methods.In addition,recent applications of DIHM technique for label-free identification and digital tracking of various motile biological cells,including human blood cells,spermatozoa,diseased cells,and unicellular microorganisms,are thoroughly examined.Leveraging artificial intelligence has significantly enhanced both the speed and accuracy of digital image processing for cell tracking and identification.The quantitative data on cell morphology and dynamics captured by DIHM can effectively elucidate the underlying mechanisms governing various microbial behaviors and contribute to the accumulation of diagnostic databases and the development of clinical treatments.
基金supported by the National Key Research and Development Program of China(Grant Nos.2022YFA1403300 and 2019YFA0308404)the National Natural Science Foundation of China(Grant Nos.11427902,11991060,12074075,12474165,12274084,and 12241402)+5 种基金Shanghai Municipal Science and Technology Major Project(Grant No.2019SHZDZX01)Shanghai Municipal Natural Science Foundation(Grant No.22ZR1407400)Innovation Program for Quantum Science and Technology(Grant No.2024ZD0300104)Innovation Program of Shanghai Municipal Education Commission(Grant No.2023ZKZD03)Science and Technology Commission of Shanghai Municipality(Grant No.20JC1415900)China Postdoctoral Science Foundation(Grant No.KLH1512149).
摘要Recently,charged solitons have been found in a two-dimensional CoCl2/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl2 using scanning tunneling microscopy(STM)and atomic force microscopy(AFM).Moreover,we study the electrical properties of the charged solitons at zero electric field by measuring local contact potential difference(LCPD)via Kelvin probe force microscopy(KPFM)using the Δf(V)method.The compensation voltage corresponding to the vertex of the parabola is obtained by fitting the quadratic relationship between Δf and sample bias.The results show that,without an external electric field,the solitons behave as negatively charged entities.Meanwhile,the LCPD mapping characterizes the spatial distribution of the potential at the charged solitons,which agrees well with those obtained from STM band bending measurements.
基金financially supported by the National Science Foundation of China(No.12274011)。
摘要The photocatalytic activity of a photocatalyst is significantly influenced by its microstructure.Therefore,it is crucial to understand and characterize the microstructure.Transmission electron microscopy(TEM)has been an indispensable tool that provides direct visualization of the microstructure at the atomic level.Despite numerous reviews on photocatalytic processes,there has been a lack of comprehensive summaries that focus on the critical role of TEM in photocatalytic applications.This review aims to fill this gap by highlighting the significant contributions of TEM techniques in understanding key photocatalytic processes,including light absorption,charge separation and transfer,and surface reaction.In addition,this review discusses in-situ TEM techniques for observing photocatalytic reactions and explores the challenges and future perspectives related to the application of TEM in photocatalysis.