The environment of low-altitude urban airspace is complex and variable due to numerous obstacles,non-cooperative aircraft,and birds.Unmanned Aerial Vehicles(UAVs)leveraging environmental information to achieve three-d...The environment of low-altitude urban airspace is complex and variable due to numerous obstacles,non-cooperative aircraft,and birds.Unmanned Aerial Vehicles(UAVs)leveraging environmental information to achieve three-dimension collision-free trajectory planning is the prerequisite to ensure airspace security.However,the timely information of surrounding situation is difficult to acquire by UAVs,which further brings security risks.As a mature technology leveraged in traditional civil aviation,the Automatic Dependent Surveillance-Broadcast(ADS-B)realizes continuous surveillance of the information of aircraft.Consequently,we leverage ADS-B for surveillance and information broadcasting,and divide the aerial airspace into multiple sub-airspaces to improve flight safety in UAV trajectory planning.In detail,we propose the secure Sub-airSpaces Planning(SSP)algorithm and Particle Swarm Optimization Rapidly-exploring Random Trees(PSO-RRT)algorithm for the UAV trajectory planning in law-altitude airspace.The performance of the proposed algorithm is verified by simulations and the results show that SSP reduces both the maximum number of UAVs in the sub-airspace and the length of the trajectory,and PSO-RRT reduces the cost of UAV trajectory in the sub-airspace.展开更多
In recent years,notable progress has been achieved in both the hardware and algorithms of structured illumination microscopy(SIM).Nevertheless,the advancement of three-dimensional structured illumination microscopy(3D...In recent years,notable progress has been achieved in both the hardware and algorithms of structured illumination microscopy(SIM).Nevertheless,the advancement of three-dimensional structured illumination microscopy(3DSIM)has been impeded by challenges arising from the speed and intricacy of polarization modulation.We introduce a high-speed modulation 3DSIM system,leveraging the polarizationmaintaining and modulation capabilities of a digital micromirror device(DMD)in conjunction with an electrooptic modulator.The DMD-3DSIM system yields a twofold enhancement in both lateral(133 nm)and axial(300 nm)resolution compared to wide-field imaging and can acquire a data set comprising 29 sections of 1024 pixels×1024 pixels,with 15 ms exposure time and 6.75 s per volume.The versatility of the DMD-3DSIM approach was exemplified through the imaging of various specimens,including fluorescent beads,nuclear pores,microtubules,actin filaments,and mitochondria within cells,as well as plant and animal tissues.Notably,polarized 3DSIM elucidated the orientation of actin filaments.Furthermore,the implementation of diverse deconvolution algorithms further enhances 3D resolution.The DMD-based 3DSIM system presents a rapid and reliable methodology for investigating biomedical phenomena,boasting capabilities encompassing 3D superresolution,fast temporal resolution,and polarization imaging.展开更多
This work presents a method for the three-dimensional localization of individual shallow NV center in diamond,leveraging the near-field quenching effect of a gold tip.Our experimental setup involves the use of an atom...This work presents a method for the three-dimensional localization of individual shallow NV center in diamond,leveraging the near-field quenching effect of a gold tip.Our experimental setup involves the use of an atomic force microscope to precisely move the gold tip close to the NV center,while simultaneously employing a home-made confocal microscope to monitor the fluorescence of the NV center.This approach allows for lateral super-resolution,achieving a full width at half maximum(FWHM)of 38.0 nm and a location uncertainty of 0.7 nm.Additionally,we show the potential of this method for determining the depth of the NV centers.We also attempt to determine the depth of the NV centers in combination with finite-difference time-domain(FDTD)simulations.Compared to other depth determination methods,this approach allows for simultaneous lateral and longitudinal localization of individual NV centers,and holds promise for facilitating manipulation of the local environment surrounding the NV center.展开更多
BACKGROUND Sigmoid colon cancer faces challenges due to anatomical diversity,including variable inferior mesenteric artery(IMA)branching and tumor localization complexities,which increase intraoperative risks.AIM To c...BACKGROUND Sigmoid colon cancer faces challenges due to anatomical diversity,including variable inferior mesenteric artery(IMA)branching and tumor localization complexities,which increase intraoperative risks.AIM To comprehensively evaluate the impact of three-dimensional(3D)visualization technology on enhancing surgical precision and safety,as well as optimizing perioperative outcomes in laparoscopic sigmoid cancer resection.METHODS A prospective cohort of 106 patients(January 2023 to December 2024)undergoing laparoscopic sigmoid cancer resection was divided into the 3D(n=55)group and the control(n=51)group.The 3D group underwent preoperative enhanced computed tomography reconstruction(3D Slicer 5.2.2&Mimics 19.0).3D reconstruction visualization navigation intraoperatively guided the following key steps:Tumor location,Toldt’s space dissection,IMA ligation level selection,regional lymph node dissection,and marginal artery preservation.Outcomes included operative parameters,lymph node yield,and recovery metrics.RESULTS The 3D group demonstrated a significantly shorter operative time(172.91±20.69 minutes vs 190.29±32.29 minutes;P=0.002),reduced blood loss(31.5±11.8 mL vs 44.1±23.4 mL,P=0.001),earlier postoperative flatus(2.23±0.54 days vs 2.53±0.61 days;P=0.013),shorter hospital length of stay(13.47±1.74 days vs 16.20±7.71 days;P=0.013),shorter postoperative length of stay(8.6±2.6 days vs 10.5±4.9 days;P=0.014),and earlier postoperative exhaust time(2.23±0.54 days vs 2.53±0.61 days;P=0.013).Furthermore,the 3D group exhibited a higher mean number of lymph nodes harvested(16.91±5.74 vs 14.45±5.66;P=0.030).CONCLUSION The 3D visualization technology effectively addresses sigmoid colon anatomical complexity through surgical navigation,improving procedural safety and efficiency.展开更多
Nano-scale chemical inhomogeneity in surface oxide films formed on a V-and N-containing martensite stainless steel and tempering heating induced changes are investigated by a combination of synchrotron-based hard X-ra...Nano-scale chemical inhomogeneity in surface oxide films formed on a V-and N-containing martensite stainless steel and tempering heating induced changes are investigated by a combination of synchrotron-based hard X-ray Photoelectron emission spectroscopy(HAXPES)and microscopy(HAXPEEM)as well as microscopic X-ray absorption spectroscopy(μ-XAS)techniques.The results reveal the inhomogeneity in the oxide films on the micron-sized Cr_(2)N-and VN-type particles,while the inhomogeneity on the martensite matrix phase exists due to localised formation of nano-sized tempering nitride particles at 600℃.The oxide film formed on Cr_(2)N-type particles is rich in Cr_(2)O_(3) compared with that on the martensite matrix and VN-type particles.With the increase of tempering temperature,Cr_(2)O_(3) formation is faster for the oxidation of Cr in the martensite matrix than the oxidation of Cr nitride-rich particles.展开更多
Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some int...Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some interesting structural features were found inside and outside of the melanosomes. By SEM observation, the melanosomes were ellipsoid shape, their surface was not smooth and was covered with rough substructure, 10 to 20 nm particles. By TEM, uneven structure and micro particles were observed in the melanosomes. Furthermore, three-dimensional analysis was tried by using the ultra-high voltage electron microscopy(UHVEM). Micrographs of the melanosomes were taken at various tilted angles by UHVEM, after preparing 500 nm thickness specimens stained with lead citrate. From the micrographs collected, the three-dimensional structures were reconstructed by using i-mode software. Melanin stained by lead and non stained parts was clearly observed in the reconstructed structure. Non stained parts were round, regular size, and distributed widely in the melanosomes.展开更多
We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse bra...We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse brain imaging,the brain tissue is 3D imaged under eight views in our study,by a home-built selective plane ilumination microscopy(SPIM).An output image containing complete structural infornation as well as significantly improved res olution(~4 times)are then computed based on these eight views of data,using a bead-guided multi-view registration and deconvolution.With superior imaging quality,the astrocyte and pyrarmidal neurons together with their subcellular nerve fbers can be clearly visualized and segmented.With further incuding other computational methods,this study can be potentially scaled up to map the conectome of whole mouse brain with a simple light.sheet microscope.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
Full-color imaging is essential in digital pathology for accurate tissue analysis.Utilizing advanced optical modulation and phase retrieval algorithms,Fourier ptychographic microscopy(FPM)offers a powerful solution fo...Full-color imaging is essential in digital pathology for accurate tissue analysis.Utilizing advanced optical modulation and phase retrieval algorithms,Fourier ptychographic microscopy(FPM)offers a powerful solution for high-throughput digital pathology,combining high resolution,large field of view,and extended depth of field(DOF).However,the full-color capabilities of FPM are hindered by coherent color artifacts and reduced computational efficiency,which significantly limits its practical applications.Color-transferbased FPM(CFPM)has emerged as a potential solution,theoretically reducing both acquisition and reconstruction threefold time.Yet,existing methods fall short of achieving the desired reconstruction speed and colorization quality.In this study,we report a generalized dual-color-space constrained model for FPM colorization.This model provides a mathematical framework for model-based FPM colorization,enabling a closed-form solution without the need for redundant iterative calculations.Our approach,termed generalized CFPM(gCFPM),achieves colorization within seconds for megapixel-scale images,delivering superior colorization quality in terms of both colorfulness and sharpness,along with an extended DOF.Both simulations and experiments demonstrate that gCFPM surpasses state-of-the-art methods across all evaluated criteria.Our work offers a robust and comprehensive workflow for high-throughput full-color pathological imaging using FPM platforms,laying a solid foundation for future advancements in methodology and engineering.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/Al_(2)O_(3)and CaO/SiO_(2)ratios in liquid slag promoted the dissolution of Al_(2)O_(3)-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
High-resolution transmission electron microscopy(HRTEM)promises rapid atomic-scale dynamic structure imaging.Yet,the precision limitations of aberration parameters and the challenge of eliminating aberrations in Cs-co...High-resolution transmission electron microscopy(HRTEM)promises rapid atomic-scale dynamic structure imaging.Yet,the precision limitations of aberration parameters and the challenge of eliminating aberrations in Cs-corrected transmission electron microscopy constrain resolution.A machine learning algorithm is developed to determine the aberration parameters with higher precision from small,lattice-periodic crystal images.The proposed algorithm is then validated with simulated HRTEM images of graphene and applied to the experimental images of a molybdenum disulfide(MoS_(2))monolayer with 25 variables(14 aberrations)resolved in wide ranges.Using these measured parameters,the phases of the exit-wave functions are reconstructed for each image in a focal series of MoS_(2)monolayers.The images were acquired due to the unexpected movement of the specimen holder.Four-dimensional data extraction reveals time-varying atomic structures and ripple.In particular,the atomic evolution of the sulfur-vacancy point and line defects,as well as the edge structure near the amorphous,is visualized as the resolution has been improved from about 1.75?to 0.9 A.This method can help salvage important transmission electron microscope images and is beneficial for the images obtained from electron microscopes with average stability.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
Enamel demineralization often occurs in the early stage of dental caries.Studying the microscopic mechanism of enamel demineralization is essential to prevent and treat dental caries.Terahertz(THz)technolo⁃gy,especial...Enamel demineralization often occurs in the early stage of dental caries.Studying the microscopic mechanism of enamel demineralization is essential to prevent and treat dental caries.Terahertz(THz)technolo⁃gy,especially continuous wave(CW)THz near-field scanning microscopy(THz-SNOM)with its nanoscale reso⁃lution,can be promising in biomedical imaging.In addition,compared with traditional THz time-domain spec⁃troscopy(TDS),portable solid-state source as the emission has higher power and SNR,lower cost,and can ob⁃tain more precise imaging.In this study,we employ CW THz-SNOM to further break the resolution limitations of conventional THz imaging techniques and successfully achieve the near-field imaging of demineralized enamel at the nanoscale.We keenly observe that the near-field signal of the enamel significantly lowers as demineralization deepens,mainly due to the decrease in permittivity.This new approach offers valuable insights into the micro⁃scopic processes of enamel demineralization,laying the foundation for further research and treatment.展开更多
Stochastic optical reconstruction microscopy(STORM),as a typical technique of single-molecule localization microscopy(SMLM),has overcome the diffraction limit by randomly switching fluorophores between fluorescent and...Stochastic optical reconstruction microscopy(STORM),as a typical technique of single-molecule localization microscopy(SMLM),has overcome the diffraction limit by randomly switching fluorophores between fluorescent and dark states,allowing for the precise localization of isolated emission patterns and the super-resolution reconstruction from millions of localized positions of single fluorophores.A critical factor influencing localization precision is the photo-switching behavior of fluorophores,which is affected by the imaging buffer.The imaging buffer typically comprises oxygen scavengers,photo-switching reagents,and refractive index regulators.Oxygen scavengers help prevent photobleaching,photo-switching reagents assist in facilitating the conversion of fluorophores,and refractive index regulators are used to adjust the refractive index of the solution.The synergistic interaction of these components promotes stable blinking of fluorophores,reduces irreversible photobleaching,and thereby ensures high-quality super-resolution imaging.This review provides a comprehensive overview of the essential compositions and functionalities of imaging buffers used in STORM,serving as a valuable resource for researchers seeking to select appropriate imaging buffers for their experiments.展开更多
Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period...Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period,exploring the full AD pathogenesis and seeking the optimal therapeutic solution have become critical and imperative.This allows researchers to intervene,delay,and potentially prevent AD progression.Several clinical imaging methods are utilized routinely to diagnose and monitor AD,such as magnetic resonance imaging(MRI),functional magnetic resonance imaging(fMRI),positron emission tomography(PET),and single photon emission computed tomography(SPECT).Nevertheless,due to their intrinsic drawbacks and restrictions,such as radiation concerns,high cost,long acquisition time,and low spatial resolution,their applications in AD research are limited,especially at the cellular and molecular levels.In contrast,optical microscopic imaging methods overcome these limitations,offering researchers a variety of approaches with distinct advantages to explore AD pathology on diverse models.In this review,we provide a comprehensive overview of commonly utilized optical microscopic imaging techniques in AD research and introduce their contributions to image amyloid beta(Aβ)species.These techniques include fluorescence microscopy(FM),confocal microscopy(CM),two-photon fluorescence microscopy(TPFM),super-resolution microscopy(SRM),expansion microscopy(ExM),and light-sheet fluorescence microscopy(LSFM).In addition,we introduce some related topics,such as the development of near-infrared(NIR)Aβprobes,the Aβplaque hypothesis,and Aβoligomer hypothesis,and the roles of microglia and astrocytes in AD progression.We believe optical microscopic imaging methods continue to play an indispensable role in deciphering the full pathogenesis of AD and advancing therapeutic strategies.展开更多
Recently,charged solitons have been found in a two-dimensional CoCl_(2)/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl_(2) using scanning ...Recently,charged solitons have been found in a two-dimensional CoCl_(2)/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl_(2) 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
TIRF microscopy has provided a means to view mobile granules within 100 nm in size in two dimensions.However quantitative analysis of the position and motion of those granules requires an appropriate tracking method.I...TIRF microscopy has provided a means to view mobile granules within 100 nm in size in two dimensions.However quantitative analysis of the position and motion of those granules requires an appropriate tracking method.In this paper,we present a new tracking algorithm combined with the unique features of TIRF.Firstly a fluorescence correction procedure was processed to solve the problem of fluorescence bleaching over time.Mobile granules were then segmented from a time-lapse image stack by an adaptive background subtraction method.Kalman filter was introduced to estimate and track the granules that allowed reducing searching range and hence greater reliability in tracking process.After the tracked granules were located in x-y plane,the z-position was indirectly inferred from the changes in their intensities.In the experiments the algorithm was applied in tracking GLUT4 vesicles in living adipose cells.The results indicate that the algorithm has achieved robust estimation and tracking of the vesicles in three dimensions.展开更多
Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and mol...Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and molecular changes remain unclear.This study investigated platelet-derived growth factor receptor beta-positive(Pdgfrb+)pericyte dynamics and reprogramming in GBM vasculature.Methods:We generated GL261-Luc and GL261-CFP glioblastoma cells via lentiviral transduction and established two transgenic models.(1)For pericyte labeling,Ai14 reporter mice was crossed with PDGFRβ-P2A-CreERT2mice for td Tomato-specific lineage tracing(PT mice).(2)For conditional ablation,we generated inducible Pdgfrb-expressing cell ablation models(PT mice was crossed with ROSA-DTA mice).An intravital imaging platform(FITC-dextran/CFP/td Tomato+two-photon microscopy)tracked pericytes,vessels,and tumor cells,while FACSsorted Pdgfrb+cells from GBM and normal brain were analyzed by LC-MS/MS proteomics.Results:Cre-mediated ablation of Pdgfrb-expressing cells revealed stage-dependent effects on GBM growth:early ablation inhibited progression while late ablation promoted it.Pericytes undergo dual spatial reorganization in GBM:regional enrichment with pre-sprouting accumulation at the tumor-brain interface,and focal positioning with preferential localization at vascular branch points.Concurrently,GBM vasculature displayed simplified branching,dilation,and pericyte remodeling(shorter processes,higher density).Proteomics revealed 1426 altered proteins,with upregulated proliferation pathways(e.g.,matrix metallopeptidase 14[Mmp14],lysyl oxidase like 2[Loxl2])and downregulated homeostasis functions(e.g.,transforming growth factor beta 1[Tgfb1]),validated by scRNA-seq in human GBM.Conclusions:This study demonstrates that during early GBM progression,pericytes actively drive tumor angiogenesis through molecular reprogramming toward proliferative and pro-angiogenic phenotypes,with the integrated imaging-proteomics framework revealing potential therapeutic targets for disrupting pericyte-mediated vascular remodeling.展开更多
Maintaining the s-polarization state of laser beams is important to achieve high modulation depth in a laser-interference-based super-resolution structured illumination microscope(SR-SIM).However,the imperfect optical...Maintaining the s-polarization state of laser beams is important to achieve high modulation depth in a laser-interference-based super-resolution structured illumination microscope(SR-SIM).However,the imperfect optical components can depolarize the laser beams hence degenerating the modulation depth.Here,we first presented a direct measurement method designed to estimate the modulation depth more precisely by shifting illumination patterns with equal phase steps.This measurement method greatly reduces the dependence of modulation depths on the samples,and then developed a polarization optimization method to achieve high modulation depth at all orientations by actively and quantitatively compensating for the additional phase difference using a combination of waveplate and a liquid crystal variable retarder(LCVR).Experimental results demonstrate that our method can achieve illumination patterns with modulation depth higher than 0.94 at three orientations with only one LCVR voltage,which enables isotropic resolution improvement.展开更多
Holographic microscopy has emerged as a vital tool in biomedicine,enabling visualization of microscopic morphological features of tissues and cells in a label-free manner.Recently,deep learning(DL)-based image reconst...Holographic microscopy has emerged as a vital tool in biomedicine,enabling visualization of microscopic morphological features of tissues and cells in a label-free manner.Recently,deep learning(DL)-based image reconstruction models have demonstrated state-of-the-art performance in holographic image reconstruction.However,their utility in practice is still severely limited,as conventional training schemes could not properly handle out-of-distribution data.Here,we leverage backpropagation operation and reparameterization of the forward propagator to enable an adaptable image reconstruction model for histopathologic inspection.Only given with a training dataset of rectum tissue images captured from a single imaging configuration,our scheme consistently shows high reconstruction performance even with the input hologram of diverse tissue types at different pathological states captured under various imaging configurations.Using the proposed adaptation technique,we show that the diagnostic features of cancerous colorectal tissues,such as dirty necrosis,captured with 5×magnification and a numerical aperture(NA)of 0.1,can be reconstructed with high accuracy,whereas a given training dataset is strictly confined to normal rectum tissues acquired under the imaging configuration of 20×magnification and an NA of 0.4.Our results suggest that the DL-based image reconstruction approaches,with sophisticated adaptation techniques,could offer an extensively generalizable solution for inverse mapping problems in imaging.展开更多
基金supported by the National Key R&D Program of China(No.2022YFB3104502)the National Natural Science Foundation of China(No.62301251)+2 种基金the Natural Science Foundation of Jiangsu Province of China under Project(No.BK20220883)the open research fund of National Mobile Communications Research Laboratory,Southeast University,China(No.2024D04)the Young Elite Scientists Sponsorship Program by CAST(No.2023QNRC001).
文摘The environment of low-altitude urban airspace is complex and variable due to numerous obstacles,non-cooperative aircraft,and birds.Unmanned Aerial Vehicles(UAVs)leveraging environmental information to achieve three-dimension collision-free trajectory planning is the prerequisite to ensure airspace security.However,the timely information of surrounding situation is difficult to acquire by UAVs,which further brings security risks.As a mature technology leveraged in traditional civil aviation,the Automatic Dependent Surveillance-Broadcast(ADS-B)realizes continuous surveillance of the information of aircraft.Consequently,we leverage ADS-B for surveillance and information broadcasting,and divide the aerial airspace into multiple sub-airspaces to improve flight safety in UAV trajectory planning.In detail,we propose the secure Sub-airSpaces Planning(SSP)algorithm and Particle Swarm Optimization Rapidly-exploring Random Trees(PSO-RRT)algorithm for the UAV trajectory planning in law-altitude airspace.The performance of the proposed algorithm is verified by simulations and the results show that SSP reduces both the maximum number of UAVs in the sub-airspace and the length of the trajectory,and PSO-RRT reduces the cost of UAV trajectory in the sub-airspace.
基金supported by the National Key R&D Program of China (Award No.2022YFC3401100)the National Natural Science Foundation of China。
文摘In recent years,notable progress has been achieved in both the hardware and algorithms of structured illumination microscopy(SIM).Nevertheless,the advancement of three-dimensional structured illumination microscopy(3DSIM)has been impeded by challenges arising from the speed and intricacy of polarization modulation.We introduce a high-speed modulation 3DSIM system,leveraging the polarizationmaintaining and modulation capabilities of a digital micromirror device(DMD)in conjunction with an electrooptic modulator.The DMD-3DSIM system yields a twofold enhancement in both lateral(133 nm)and axial(300 nm)resolution compared to wide-field imaging and can acquire a data set comprising 29 sections of 1024 pixels×1024 pixels,with 15 ms exposure time and 6.75 s per volume.The versatility of the DMD-3DSIM approach was exemplified through the imaging of various specimens,including fluorescent beads,nuclear pores,microtubules,actin filaments,and mitochondria within cells,as well as plant and animal tissues.Notably,polarized 3DSIM elucidated the orientation of actin filaments.Furthermore,the implementation of diverse deconvolution algorithms further enhances 3D resolution.The DMD-based 3DSIM system presents a rapid and reliable methodology for investigating biomedical phenomena,boasting capabilities encompassing 3D superresolution,fast temporal resolution,and polarization imaging.
基金supported by the National Natural Science Foundation of China(T2325023,92265204,12104447)the National Key R&D Program of China(2023YFF0718400)+1 种基金the Innovation Program for Quantum Science and Technology(2021ZD0302200)the Fundamental Research Funds for the Central Universities。
文摘This work presents a method for the three-dimensional localization of individual shallow NV center in diamond,leveraging the near-field quenching effect of a gold tip.Our experimental setup involves the use of an atomic force microscope to precisely move the gold tip close to the NV center,while simultaneously employing a home-made confocal microscope to monitor the fluorescence of the NV center.This approach allows for lateral super-resolution,achieving a full width at half maximum(FWHM)of 38.0 nm and a location uncertainty of 0.7 nm.Additionally,we show the potential of this method for determining the depth of the NV centers.We also attempt to determine the depth of the NV centers in combination with finite-difference time-domain(FDTD)simulations.Compared to other depth determination methods,this approach allows for simultaneous lateral and longitudinal localization of individual NV centers,and holds promise for facilitating manipulation of the local environment surrounding the NV center.
基金Supported by the Health Commission of Fuyang City,Anhui,China,No.FY2023-45Fuyang Municipal Science and Technology Bureau,Anhui,China,No.FK20245505+1 种基金Anhui Provincial Health Commission,No.AHWJ2023Baa20164Bengbu Medical University,No.2023byzd215.
文摘BACKGROUND Sigmoid colon cancer faces challenges due to anatomical diversity,including variable inferior mesenteric artery(IMA)branching and tumor localization complexities,which increase intraoperative risks.AIM To comprehensively evaluate the impact of three-dimensional(3D)visualization technology on enhancing surgical precision and safety,as well as optimizing perioperative outcomes in laparoscopic sigmoid cancer resection.METHODS A prospective cohort of 106 patients(January 2023 to December 2024)undergoing laparoscopic sigmoid cancer resection was divided into the 3D(n=55)group and the control(n=51)group.The 3D group underwent preoperative enhanced computed tomography reconstruction(3D Slicer 5.2.2&Mimics 19.0).3D reconstruction visualization navigation intraoperatively guided the following key steps:Tumor location,Toldt’s space dissection,IMA ligation level selection,regional lymph node dissection,and marginal artery preservation.Outcomes included operative parameters,lymph node yield,and recovery metrics.RESULTS The 3D group demonstrated a significantly shorter operative time(172.91±20.69 minutes vs 190.29±32.29 minutes;P=0.002),reduced blood loss(31.5±11.8 mL vs 44.1±23.4 mL,P=0.001),earlier postoperative flatus(2.23±0.54 days vs 2.53±0.61 days;P=0.013),shorter hospital length of stay(13.47±1.74 days vs 16.20±7.71 days;P=0.013),shorter postoperative length of stay(8.6±2.6 days vs 10.5±4.9 days;P=0.014),and earlier postoperative exhaust time(2.23±0.54 days vs 2.53±0.61 days;P=0.013).Furthermore,the 3D group exhibited a higher mean number of lymph nodes harvested(16.91±5.74 vs 14.45±5.66;P=0.030).CONCLUSION The 3D visualization technology effectively addresses sigmoid colon anatomical complexity through surgical navigation,improving procedural safety and efficiency.
基金supported by the Vinnova(project number 2020-03778)supported by the Swedish Research Council(Vetenskapsradet,project number 2021-04157).
文摘Nano-scale chemical inhomogeneity in surface oxide films formed on a V-and N-containing martensite stainless steel and tempering heating induced changes are investigated by a combination of synchrotron-based hard X-ray Photoelectron emission spectroscopy(HAXPES)and microscopy(HAXPEEM)as well as microscopic X-ray absorption spectroscopy(μ-XAS)techniques.The results reveal the inhomogeneity in the oxide films on the micron-sized Cr_(2)N-and VN-type particles,while the inhomogeneity on the martensite matrix phase exists due to localised formation of nano-sized tempering nitride particles at 600℃.The oxide film formed on Cr_(2)N-type particles is rich in Cr_(2)O_(3) compared with that on the martensite matrix and VN-type particles.With the increase of tempering temperature,Cr_(2)O_(3) formation is faster for the oxidation of Cr in the martensite matrix than the oxidation of Cr nitride-rich particles.
文摘Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some interesting structural features were found inside and outside of the melanosomes. By SEM observation, the melanosomes were ellipsoid shape, their surface was not smooth and was covered with rough substructure, 10 to 20 nm particles. By TEM, uneven structure and micro particles were observed in the melanosomes. Furthermore, three-dimensional analysis was tried by using the ultra-high voltage electron microscopy(UHVEM). Micrographs of the melanosomes were taken at various tilted angles by UHVEM, after preparing 500 nm thickness specimens stained with lead citrate. From the micrographs collected, the three-dimensional structures were reconstructed by using i-mode software. Melanin stained by lead and non stained parts was clearly observed in the reconstructed structure. Non stained parts were round, regular size, and distributed widely in the melanosomes.
基金funding support from 1000 Youth Talents Plan of China (P.F.)Fundamental Research Program of Shenzhen (P.F.,JCYJ20160429182424047)+1 种基金National Science Foundation of China (NSFC31571002,D.Z)Graduates'Innovation Fund of Huazhong University of Science and Technology (5003182004).
文摘We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse brain imaging,the brain tissue is 3D imaged under eight views in our study,by a home-built selective plane ilumination microscopy(SPIM).An output image containing complete structural infornation as well as significantly improved res olution(~4 times)are then computed based on these eight views of data,using a bead-guided multi-view registration and deconvolution.With superior imaging quality,the astrocyte and pyrarmidal neurons together with their subcellular nerve fbers can be clearly visualized and segmented.With further incuding other computational methods,this study can be potentially scaled up to map the conectome of whole mouse brain with a simple light.sheet microscope.
基金supported by the Project of Health Committee of Hunan Province(D202304128868),China.
文摘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 Natural Science Foundation of China(Grant Nos.12104500 and 82430062)the Key Research and Development Projects of Shaanxi Province(Grant No.2023-YBSF-263),the Shenzhen Engineering Research Centre(Grant No.XMHT20230115004)the Shenzhen Science and Technology Innovation Commission(Grant No.KCXFZ20201221173207022).
文摘Full-color imaging is essential in digital pathology for accurate tissue analysis.Utilizing advanced optical modulation and phase retrieval algorithms,Fourier ptychographic microscopy(FPM)offers a powerful solution for high-throughput digital pathology,combining high resolution,large field of view,and extended depth of field(DOF).However,the full-color capabilities of FPM are hindered by coherent color artifacts and reduced computational efficiency,which significantly limits its practical applications.Color-transferbased FPM(CFPM)has emerged as a potential solution,theoretically reducing both acquisition and reconstruction threefold time.Yet,existing methods fall short of achieving the desired reconstruction speed and colorization quality.In this study,we report a generalized dual-color-space constrained model for FPM colorization.This model provides a mathematical framework for model-based FPM colorization,enabling a closed-form solution without the need for redundant iterative calculations.Our approach,termed generalized CFPM(gCFPM),achieves colorization within seconds for megapixel-scale images,delivering superior colorization quality in terms of both colorfulness and sharpness,along with an extended DOF.Both simulations and experiments demonstrate that gCFPM surpasses state-of-the-art methods across all evaluated criteria.Our work offers a robust and comprehensive workflow for high-throughput full-color pathological imaging using FPM platforms,laying a solid foundation for future advancements in methodology and engineering.
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.
文摘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/Al_(2)O_(3)and CaO/SiO_(2)ratios in liquid slag promoted the dissolution of Al_(2)O_(3)-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financial support from the National Natural Science Foundation of China(Grant No.61971201)。
文摘High-resolution transmission electron microscopy(HRTEM)promises rapid atomic-scale dynamic structure imaging.Yet,the precision limitations of aberration parameters and the challenge of eliminating aberrations in Cs-corrected transmission electron microscopy constrain resolution.A machine learning algorithm is developed to determine the aberration parameters with higher precision from small,lattice-periodic crystal images.The proposed algorithm is then validated with simulated HRTEM images of graphene and applied to the experimental images of a molybdenum disulfide(MoS_(2))monolayer with 25 variables(14 aberrations)resolved in wide ranges.Using these measured parameters,the phases of the exit-wave functions are reconstructed for each image in a focal series of MoS_(2)monolayers.The images were acquired due to the unexpected movement of the specimen holder.Four-dimensional data extraction reveals time-varying atomic structures and ripple.In particular,the atomic evolution of the sulfur-vacancy point and line defects,as well as the edge structure near the amorphous,is visualized as the resolution has been improved from about 1.75?to 0.9 A.This method can help salvage important transmission electron microscope images and is beneficial for the images obtained from electron microscopes with average stability.
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)。
文摘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(61988102,62401113,92463308)。
文摘Enamel demineralization often occurs in the early stage of dental caries.Studying the microscopic mechanism of enamel demineralization is essential to prevent and treat dental caries.Terahertz(THz)technolo⁃gy,especially continuous wave(CW)THz near-field scanning microscopy(THz-SNOM)with its nanoscale reso⁃lution,can be promising in biomedical imaging.In addition,compared with traditional THz time-domain spec⁃troscopy(TDS),portable solid-state source as the emission has higher power and SNR,lower cost,and can ob⁃tain more precise imaging.In this study,we employ CW THz-SNOM to further break the resolution limitations of conventional THz imaging techniques and successfully achieve the near-field imaging of demineralized enamel at the nanoscale.We keenly observe that the near-field signal of the enamel significantly lowers as demineralization deepens,mainly due to the decrease in permittivity.This new approach offers valuable insights into the micro⁃scopic processes of enamel demineralization,laying the foundation for further research and treatment.
基金funded by the National Natural Science Foundation of China(No.62305041)the Natural Science Foundation of Liaoning Province(No.2023-MS-103)。
文摘Stochastic optical reconstruction microscopy(STORM),as a typical technique of single-molecule localization microscopy(SMLM),has overcome the diffraction limit by randomly switching fluorophores between fluorescent and dark states,allowing for the precise localization of isolated emission patterns and the super-resolution reconstruction from millions of localized positions of single fluorophores.A critical factor influencing localization precision is the photo-switching behavior of fluorophores,which is affected by the imaging buffer.The imaging buffer typically comprises oxygen scavengers,photo-switching reagents,and refractive index regulators.Oxygen scavengers help prevent photobleaching,photo-switching reagents assist in facilitating the conversion of fluorophores,and refractive index regulators are used to adjust the refractive index of the solution.The synergistic interaction of these components promotes stable blinking of fluorophores,reduces irreversible photobleaching,and thereby ensures high-quality super-resolution imaging.This review provides a comprehensive overview of the essential compositions and functionalities of imaging buffers used in STORM,serving as a valuable resource for researchers seeking to select appropriate imaging buffers for their experiments.
基金supported by NIH(R01AG055413),(R01AG085562),(R21AG059134),(R21AG078749),and(S10OD028609)awards(C.R.).NIH Office of the Director,National Institute on Aging.
文摘Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period,exploring the full AD pathogenesis and seeking the optimal therapeutic solution have become critical and imperative.This allows researchers to intervene,delay,and potentially prevent AD progression.Several clinical imaging methods are utilized routinely to diagnose and monitor AD,such as magnetic resonance imaging(MRI),functional magnetic resonance imaging(fMRI),positron emission tomography(PET),and single photon emission computed tomography(SPECT).Nevertheless,due to their intrinsic drawbacks and restrictions,such as radiation concerns,high cost,long acquisition time,and low spatial resolution,their applications in AD research are limited,especially at the cellular and molecular levels.In contrast,optical microscopic imaging methods overcome these limitations,offering researchers a variety of approaches with distinct advantages to explore AD pathology on diverse models.In this review,we provide a comprehensive overview of commonly utilized optical microscopic imaging techniques in AD research and introduce their contributions to image amyloid beta(Aβ)species.These techniques include fluorescence microscopy(FM),confocal microscopy(CM),two-photon fluorescence microscopy(TPFM),super-resolution microscopy(SRM),expansion microscopy(ExM),and light-sheet fluorescence microscopy(LSFM).In addition,we introduce some related topics,such as the development of near-infrared(NIR)Aβprobes,the Aβplaque hypothesis,and Aβoligomer hypothesis,and the roles of microglia and astrocytes in AD progression.We believe optical microscopic imaging methods continue to play an indispensable role in deciphering the full pathogenesis of AD and advancing therapeutic strategies.
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).
文摘Recently,charged solitons have been found in a two-dimensional CoCl_(2)/HOPG system,whose microscopic nature remains to be elusive.In this work,we investigate the charged solitons in monolayer CoCl_(2) 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Project supported by the National Natural Science Foundation ofChina (No. 30770596)the Key Laboratory for Biomedical En-gineering of Ministry of Education of China
文摘TIRF microscopy has provided a means to view mobile granules within 100 nm in size in two dimensions.However quantitative analysis of the position and motion of those granules requires an appropriate tracking method.In this paper,we present a new tracking algorithm combined with the unique features of TIRF.Firstly a fluorescence correction procedure was processed to solve the problem of fluorescence bleaching over time.Mobile granules were then segmented from a time-lapse image stack by an adaptive background subtraction method.Kalman filter was introduced to estimate and track the granules that allowed reducing searching range and hence greater reliability in tracking process.After the tracked granules were located in x-y plane,the z-position was indirectly inferred from the changes in their intensities.In the experiments the algorithm was applied in tracking GLUT4 vesicles in living adipose cells.The results indicate that the algorithm has achieved robust estimation and tracking of the vesicles in three dimensions.
基金The National Key Research and Development Program of China,Grant/Award Number:2022YFF0710700Nonprofit Central Research Institute Fund of Chinese Academy of Medical Sciences,Grant/Award Number:2023-PT180-01。
文摘Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and molecular changes remain unclear.This study investigated platelet-derived growth factor receptor beta-positive(Pdgfrb+)pericyte dynamics and reprogramming in GBM vasculature.Methods:We generated GL261-Luc and GL261-CFP glioblastoma cells via lentiviral transduction and established two transgenic models.(1)For pericyte labeling,Ai14 reporter mice was crossed with PDGFRβ-P2A-CreERT2mice for td Tomato-specific lineage tracing(PT mice).(2)For conditional ablation,we generated inducible Pdgfrb-expressing cell ablation models(PT mice was crossed with ROSA-DTA mice).An intravital imaging platform(FITC-dextran/CFP/td Tomato+two-photon microscopy)tracked pericytes,vessels,and tumor cells,while FACSsorted Pdgfrb+cells from GBM and normal brain were analyzed by LC-MS/MS proteomics.Results:Cre-mediated ablation of Pdgfrb-expressing cells revealed stage-dependent effects on GBM growth:early ablation inhibited progression while late ablation promoted it.Pericytes undergo dual spatial reorganization in GBM:regional enrichment with pre-sprouting accumulation at the tumor-brain interface,and focal positioning with preferential localization at vascular branch points.Concurrently,GBM vasculature displayed simplified branching,dilation,and pericyte remodeling(shorter processes,higher density).Proteomics revealed 1426 altered proteins,with upregulated proliferation pathways(e.g.,matrix metallopeptidase 14[Mmp14],lysyl oxidase like 2[Loxl2])and downregulated homeostasis functions(e.g.,transforming growth factor beta 1[Tgfb1]),validated by scRNA-seq in human GBM.Conclusions:This study demonstrates that during early GBM progression,pericytes actively drive tumor angiogenesis through molecular reprogramming toward proliferative and pro-angiogenic phenotypes,with the integrated imaging-proteomics framework revealing potential therapeutic targets for disrupting pericyte-mediated vascular remodeling.
基金supported by the National Natural Science Foundation of China[Grant Nos.62205367 and 62141506]the Suzhou Basic Research Pilot Project[Grant Nos.SSD2023006 and SJC2021013]the National Key Research and Development Program of China[Grant No.2023YFF1205700].
文摘Maintaining the s-polarization state of laser beams is important to achieve high modulation depth in a laser-interference-based super-resolution structured illumination microscope(SR-SIM).However,the imperfect optical components can depolarize the laser beams hence degenerating the modulation depth.Here,we first presented a direct measurement method designed to estimate the modulation depth more precisely by shifting illumination patterns with equal phase steps.This measurement method greatly reduces the dependence of modulation depths on the samples,and then developed a polarization optimization method to achieve high modulation depth at all orientations by actively and quantitatively compensating for the additional phase difference using a combination of waveplate and a liquid crystal variable retarder(LCVR).Experimental results demonstrate that our method can achieve illumination patterns with modulation depth higher than 0.94 at three orientations with only one LCVR voltage,which enables isotropic resolution improvement.
基金supported by the Samsung Research Funding and Incubation Center of Samsung Electronics(Grant No.SRFC-IT2002-03)the Samsung Electronics Co.,Ltd.(Grant No.IO220908-02403-01)+2 种基金the National Research Foundation of Korea(NRF)grant funded by the Korean government(MSIT)(Grant Nos.NRF-RS-2021-NR060086 and NRF-RS-2023-00251628)the Bio&Medical Technology Development Program of the National Research Foundation funded by the Korean government(MSIT)(Grant No RS-2024-00397673)the KAIST-CERAGEM Next Generation Healthcare Research Center.
文摘Holographic microscopy has emerged as a vital tool in biomedicine,enabling visualization of microscopic morphological features of tissues and cells in a label-free manner.Recently,deep learning(DL)-based image reconstruction models have demonstrated state-of-the-art performance in holographic image reconstruction.However,their utility in practice is still severely limited,as conventional training schemes could not properly handle out-of-distribution data.Here,we leverage backpropagation operation and reparameterization of the forward propagator to enable an adaptable image reconstruction model for histopathologic inspection.Only given with a training dataset of rectum tissue images captured from a single imaging configuration,our scheme consistently shows high reconstruction performance even with the input hologram of diverse tissue types at different pathological states captured under various imaging configurations.Using the proposed adaptation technique,we show that the diagnostic features of cancerous colorectal tissues,such as dirty necrosis,captured with 5×magnification and a numerical aperture(NA)of 0.1,can be reconstructed with high accuracy,whereas a given training dataset is strictly confined to normal rectum tissues acquired under the imaging configuration of 20×magnification and an NA of 0.4.Our results suggest that the DL-based image reconstruction approaches,with sophisticated adaptation techniques,could offer an extensively generalizable solution for inverse mapping problems in imaging.
