A new scheme of super-resolution optical fluctuation imaging(SOFI)is proposed to broaden its application in the high-order cumulant reconstruction by optimizing blinking characteristics,eliminating noise in raw data a...A new scheme of super-resolution optical fluctuation imaging(SOFI)is proposed to broaden its application in the high-order cumulant reconstruction by optimizing blinking characteristics,eliminating noise in raw data and applying multi-resolution analysis in cumulant reconstruction.A motor-driven rotating mask optical modulation system is designed to adjust the excitation lightfield and allows for fast deployment.Active-modulated fluorescence fluctuation superresolution microscopy with multi-resolution analysis(AMF-MRA-SOFI)demonstrates enhanced resolution ability and reconstruction quality in experiments performed on sample of conventional dyes,achieving a resolution of 100 nm in the fourth order compared to conventional SOFI reconstruction.Furthermore,our approach combining expansion super-resolution achieved a resolution at-57 nm.展开更多
Blood cells are the most integral part of the body,which are made up of erythrocytes,platelets and white blood cells.The examination of subcellular structures and proteins within blood cells at the nanoscale can provi...Blood cells are the most integral part of the body,which are made up of erythrocytes,platelets and white blood cells.The examination of subcellular structures and proteins within blood cells at the nanoscale can provide valuable insights into the health status of an individual,accurate diagnosis,and efficient treatment strategies for diseases.Super-resolution microscopy(SRM)has recently emerged as a cutting-edge tool for the study of blood cells,providing numerous advantages over traditional methods for examining subcellular structures and proteins.In this paper,we focus on outlining the fundamental principles of various SRM techniques and their applications in both normal and diseased states of blood cells.Furthermore,future prospects of SRM techniques in the analysis of blood cells are also discussed.展开更多
Optical microscopy promises researchers to soe most tiny substances directly.However,the resolution of conventional microscopy is resticted by the diffraction limit.This makes it a challenge to observe subcellular pro...Optical microscopy promises researchers to soe most tiny substances directly.However,the resolution of conventional microscopy is resticted by the diffraction limit.This makes it a challenge to observe subcellular processes happened in nanoscale.The development of super-resolution microscopy provides a solution to this challenge.Here,we briefly review several commonly used super-resolution techniques,explicating their basic principles and applications in biological science,especially in neuroscience.In addition,characteristics and limitations of each techrique are compared to provide a guidance for biologists to choose the most suitable tool.展开更多
Image scanning microscopy based on pixel reassignment can improve the confocal resolution limit without losing the image signal-to-noise ratio(SNR)greatly[C.J.R.Sheppard,"Super resolution in confocal imaging,&quo...Image scanning microscopy based on pixel reassignment can improve the confocal resolution limit without losing the image signal-to-noise ratio(SNR)greatly[C.J.R.Sheppard,"Super resolution in confocal imaging,"Optik 80(2)53-54(1988).C.B.Miller,E.Jorg,"Image scanning microscopy,"Phys.Reu.Lett.104(19)198101(2010).C.J.R.Sheppard,s.B.Mehta,R Heintzmann,"Superresolution by image scanning microscopy using pixel reassignment,"Opt.Lett.38(15)28892892(2013)].Here,we use a tailor-made optical fiber and 19 avalanche pho-todiodes(APDs)as parallel detectors to upgrade our existing confocal microscopy,termed as parallel-detection super resolution(PDSR)microscopy.In order to obtain the correct shift value,we use the normalized 2D cross correlation to calculate the shifting value of each image.We characterized our system performance by imaging fuorescence beads and applied this system to observing the 3D structure of biological specimen.展开更多
Remote sensing image super-resolution technology is pivotal for enhancing image quality in critical applications including environmental monitoring,urban planning,and disaster assessment.However,traditional methods ex...Remote sensing image super-resolution technology is pivotal for enhancing image quality in critical applications including environmental monitoring,urban planning,and disaster assessment.However,traditional methods exhibit deficiencies in detail recovery and noise suppression,particularly when processing complex landscapes(e.g.,forests,farmlands),leading to artifacts and spectral distortions that limit practical utility.To address this,we propose an enhanced Super-Resolution Generative Adversarial Network(SRGAN)framework featuring three key innovations:(1)Replacement of L1/L2 loss with a robust Charbonnier loss to suppress noise while preserving edge details via adaptive gradient balancing;(2)A multi-loss joint optimization strategy dynamically weighting Charbonnier loss(β=0.5),Visual Geometry Group(VGG)perceptual loss(α=1),and adversarial loss(γ=0.1)to synergize pixel-level accuracy and perceptual quality;(3)A multi-scale residual network(MSRN)capturing cross-scale texture features(e.g.,forest canopies,mountain contours).Validated on Sentinel-2(10 m)and SPOT-6/7(2.5 m)datasets covering 904 km2 in Motuo County,Xizang,our method outperforms the SRGAN baseline(SR4RS)with Peak Signal-to-Noise Ratio(PSNR)gains of 0.29 dB and Structural Similarity Index(SSIM)improvements of 3.08%on forest imagery.Visual comparisons confirm enhanced texture continuity despite marginal Learned Perceptual Image Patch Similarity(LPIPS)increases.The method significantly improves noise robustness and edge retention in complex geomorphology,demonstrating 18%faster response in forest fire early warning and providing high-resolution support for agricultural/urban monitoring.Future work will integrate spectral constraints and lightweight architectures.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
We developed an imaging technique combining two-photon computed super-resolution microscopy and suction-based stabilization to achieve the resolution of the single-cell level and organelles in vivo.To accomplish this,...We developed an imaging technique combining two-photon computed super-resolution microscopy and suction-based stabilization to achieve the resolution of the single-cell level and organelles in vivo.To accomplish this,a conventional two-photon microscope was equipped with a 3D-printed holders,which stabilize the tissue surface within the focal plane of immersion objectives.Further computational image stabilization and noise reduction were applied,followed by superresolution radial fluctuations(SRRF)analysis,doubling image resolution,and enhancing signal-to-noise ratios for in vivo subcellular process investigation.Stabilization of<1μm was obtained by suction,and<25 nm were achieved by subsequent algorithmic image stabilization.A Mito-Dendra2 mouse model,expressing green fluorescent protein(GFP)in mitochondria,demonstrated the potential of long-term intravital subcellular imaging.In vivo mitochondrial fission and fusion,mitochondrial status migration,and the effects of alcohol consumption(modeled as an alcoholic liver disease)and berberine treatment on hepatocyte mitochondrial dynamics are directly observed intravitally.Suction-based stabilization in two-photon intravital imaging,coupled with computational super-resolution holds promise for advancing in vivo subcellular imaging studies.展开更多
The broad applicability of super-resolution microscopy has been widely demonstrated in various areas and disciplines. The optimization and improvement of algorithms used in super-resolution microscopy are of great imp...The broad applicability of super-resolution microscopy has been widely demonstrated in various areas and disciplines. The optimization and improvement of algorithms used in super-resolution microscopy are of great importance for achieving optimal quality of super-resolution imaging. In this review, we comprehensively discuss the computational methods in different types of super-resolution microscopy, including deconvolution microscopy, polarization-based super-resolution microscopy, structured illumination microscopy, image scanning microscopy, super-resolution optical fluctuation imaging microscopy, single-molecule localization microscopy, Bayesian super-resolution microscopy, stimulated emission depletion microscopy, and translation microscopy. The development of novel computational methods would greatly benefit super-resolution microscopy and lead to better resolution, improved accuracy, and faster image processing.展开更多
Resolution is undoubtedly the most important parameter in optical microscopy by providing an estimation on the maximum resolving power of a certain optical microscope. For centuries, the resolution of an optical micro...Resolution is undoubtedly the most important parameter in optical microscopy by providing an estimation on the maximum resolving power of a certain optical microscope. For centuries, the resolution of an optical microscope is generally considered to be limited only by the numerical aperture of the optical system and the wavelength of light. However, since the invention and popularity of various advanced fluorescence microscopy techniques, especially super-resolution fluorescence microscopy, many new methods have been proposed for estimating the resolution, leading to confusions for researchers who need to quantify the resolution of their fluorescence microscopes. In this paper, we firstly summarize the early concepts and criteria for predicting the resolution limit of an ideal optical system. Then, we discuss some important influence factors that deteriorate the resolution of a certain fluorescence microscope. Finally, we provide methods and examples on how to measure the resolution of a fluorescence microscope from captured fluorescence images. This paper aims to answer as best as possible the theoretical and practical issues regarding the resolution estimation in fluorescence microscopy.展开更多
Various super-resolution microscopy techniques have been presented to explore fine structures of biological specimens.However,the super-resolution capability is often achieved at the expense of reducing imaging speed ...Various super-resolution microscopy techniques have been presented to explore fine structures of biological specimens.However,the super-resolution capability is often achieved at the expense of reducing imaging speed by either point scanning or multiframe computation.The contradiction between spatial resolution and imaging speed seriously hampers the observation of high-speed dynamics of fine structures.To overcome this contradiction,here we propose and demonstrate a temporal compressive super-resolution microscopy(TCSRM)technique.This technique is to merge an enhanced temporal compressive microscopy and a deep-learning-based super-resolution image reconstruction,where the enhanced temporal compressive microscopy is utilized to improve the imaging speed,and the deep-learning-based super-resolution image reconstruction is used to realize the resolution enhancement.The high-speed super-resolution imaging ability of TCSRM with a frame rate of 1200 frames per second(fps)and spatial resolution of 100 nm is experimentally demonstrated by capturing the flowing fluorescent beads in microfluidic chip.Given the outstanding imaging performance with high-speed super-resolution,TCSRM provides a desired tool for the studies of high-speed dynamical behaviors in fine structures,especially in the biomedical field.展开更多
Photon avalanche occurring in lanthanide-doped materials exhibits a giant optical nonlinear response of the emission intensity to the excitation intensity,which holds great potential in the applications of optical sen...Photon avalanche occurring in lanthanide-doped materials exhibits a giant optical nonlinear response of the emission intensity to the excitation intensity,which holds great potential in the applications of optical sensing,super-resolution imaging,quantum detection,and other techniques.However,strategies for developing photon avalanches in nanoparticles are limited,and many widely used lanthanide ions have not yet been able to generate high-efficiency avalanching emissions.A general strategy named cascade migrating photon avalanche was proposed to achieve efficient avalanching emissions with huge optical nonlinearities from a large number of emitters at the nanoscale and at room temperature.Specifically,the optical nonlinearity order of bright avalanched Tm^(3+)-emission was achieved at 63rd order by utilizing the Yb^(3+)∕Pr^(3+)-codoped nano-engine.By further incorporating a Gd^(3+)sublattice migrating network,its avalanching energy can propagate over a long distance to arouse avalanching emission with extreme optical nonlinearities up to 45th order among various emitters(Tb^(3+),Eu^(3+),Dy^(3+),Sm^(3+))in multilayered nanostructures.By achieving abundant avalanching full-spectrum emissions,it would be highly conducive to applications in various fields.For instance,our strategy demonstrated its applicability in multi-color super-resolution microscopic imaging with single-nanoparticle sensitivity and resolution up to 48 nm,utilizing a single low-power 852 nm excitation beam.展开更多
The transmembrane protein CD47,an innate immune checkpoint protein,plays a pivotal role in preventing healthy erythrocytes from immune clearance.Our study utilized stochastic optical reconstruction microscopy(STORM)an...The transmembrane protein CD47,an innate immune checkpoint protein,plays a pivotal role in preventing healthy erythrocytes from immune clearance.Our study utilized stochastic optical reconstruction microscopy(STORM)and single-molecule analysis to investigate the distribution of CD47 on the human erythrocyte membrane.Contrary to previous findings in mouse erythrocytes,we discovered that CD47 exists in randomly distributed monomers rather than in clusters across the human erythrocyte membrane.Using secondary antibody-induced crosslinking,we found that CD47 aggregates into stable clusters within minutes.By comparing these STORM results with those of the fully mobile protein CD59 and the cytoskeleton-bound membrane protein glycophorin C under similar conditions,as well as devising two-color STORM co-labeling and co-clustering experiments,we further quantitatively revealed an intermediate,self-limiting clustering behavior of CD47,elucidating its fractional(∼14%)attachment to the cytoskele-ton.Moreover,we report reductions in both the amount of CD47 and its clustering capability in aged erythrocytes,providing new insight into erythrocyte senescence.Together,the combination of STORM and secondary antibody-based crosslinking unveils the unique self-limiting clustering behavior of CD47 due to its fractional cytoskeleton attachment.展开更多
With the support by the National Natural Science Foundation of China and the Chinese Academy of Sciences,the research team led by Prof.Li Junbai(李峻柏)at the CAS Key Lab of Colloid,Interface and Thermodynamics,Instit...With the support by the National Natural Science Foundation of China and the Chinese Academy of Sciences,the research team led by Prof.Li Junbai(李峻柏)at the CAS Key Lab of Colloid,Interface and Thermodynamics,Institute of Chemistry,Chinese Academy of Sciences,revealed the distribution of proteins in the transformation of inorganic/protein hybrid crystals by super-resolution microscopy,which展开更多
The introduction of super-resolution microscopy(SRM)has significantly advanced our understanding of cellular and molecular dynamics,offering a detailed view previously beyond our reach.Implementing SRM in biophysical ...The introduction of super-resolution microscopy(SRM)has significantly advanced our understanding of cellular and molecular dynamics,offering a detailed view previously beyond our reach.Implementing SRM in biophysical research,however,presents numerous challenges.This review addresses the crucial aspects of utilizing SRM effectively,from selecting appropriate fluorophores and preparing samples to analyzing complex data sets.We explore recent technological advancements and methodological improvements that enhance the capabilities of SRM.Emphasizing the integration of SRM with other analytical methods,we aim to overcome inherent limitations and expand the scope of biological insights achievable.By providing a comprehensive guide for choosing the most suitable SRM methods based on specific research objectives,we aim to empower researchers to explore complex biological processes with enhanced precision and clarity,thereby advancing the frontiers of biophysical research.展开更多
The algorithm used for reconstruction or resolution enhancement is one of the factors affectingthe quality of super-resolution images obtained by fluorescence microscopy.Deep-learning-basedalgorithms have achieved sta...The algorithm used for reconstruction or resolution enhancement is one of the factors affectingthe quality of super-resolution images obtained by fluorescence microscopy.Deep-learning-basedalgorithms have achieved stateof-the-art performance in super-resolution fluorescence micros-copy and are becoming increasingly attractive.We firstly introduce commonly-used deep learningmodels,and then review the latest applications in terms of the net work architectures,the trainingdata and the loss functions.Additionally,we discuss the challenges and limits when using deeplearning to analyze the fluorescence microscopic data,and suggest ways to improve the reliability and robustness of deep learning applications.展开更多
The resolution of conventional optical microscopy is only -200 nm, which is becoming less and less sufficient for a variety of applications. In order to surpass the diffraction limited resolution, super-resolution mic...The resolution of conventional optical microscopy is only -200 nm, which is becoming less and less sufficient for a variety of applications. In order to surpass the diffraction limited resolution, super-resolution microscopy （SRM） has been developed to achieve a high resolution of one to tens of nanometers. The techniques involved in SRM can be assigned into two broad categories, namely ＂true＂ super-resolution techniques and ＂functional＂ super-resolution techniques. In ＂functional＂ super-resolution techniques, stochastic super-resolution microscopy （SSRM） is widely used due to its low expense, simple operation, and high resolution. The principle process in SSRM is to accumulate the coordinates of many diffraction-limited emitters （e.g., single fluorescent molecules） on the object by localizing the centroids of the point spread functions （PSF）, and then reconstruct the image of the object using these coordinates. When the diffraction-limited emitters take part in a catalytic reaction, the activity distribution and kinetic information about the catalysis by nanoparticles can be obtained by SSRM. SSRM has been applied and exhibited outstanding advantages in several fields of catalysis, such as metal nanoparticle catalysis, molecular sieve catalysis, and photocatalysis. Since SSRM is able to resolve the catalytic activity within one nanoparticle, it promises to accelerate the development and discovery of new and better catalysts. This review will present a brief introduction to SRM, and a detailed description of SSRM and its applications in nano-catalysis.展开更多
Hematologic malignancies are one of the most common malignant tumors caused by the clonal proliferation and differentiation of hematopoietic and lymphoid stem cells.The examination of bone marrow cells combined with i...Hematologic malignancies are one of the most common malignant tumors caused by the clonal proliferation and differentiation of hematopoietic and lymphoid stem cells.The examination of bone marrow cells combined with immunodeficiency typing is of great significance to the diagnostic type,treatment and prognosis of hematologic malignancies.Super-resolution fluorescence microscopy(SRM)is a special kind of optical microscopy technology,which breaks the resolution limit and was awarded the Nobel Prize in Chemistry in 2014.With the development of SRM,many related technologies have been applied to the diagnosis and treatment of clinical diseases.It was reported that a major type of SRM technique,single molecule localization microscopy(SMLM),is more sensitive than flow cytometry(FC)in detecting cell membrane antigens'expression,thus enabling better chances in detecting antigens on hematopoietic cells than traditional analytic tools.Furthermore,SRM may be applied to clinical pathology and may guide precision medicine and personalized medicine for clone hematopoietic cell diseases.In this paper,we mainly discuss the application of SRM in clone hematological malignancies.展开更多
We use the label-free microsphere-assisted microscopy to image low-contrast hexagonally close-packed polystyrene nanoparticle arrays with diameters of 300 and 250 nm.When a nanoparticle array is directly placed on a g...We use the label-free microsphere-assisted microscopy to image low-contrast hexagonally close-packed polystyrene nanoparticle arrays with diameters of 300 and 250 nm.When a nanoparticle array is directly placed on a glass slide,it cannot be distinguished.If a 30-nm-thick Ag film is deposited on the surface of a nanoparticle array,the nanoparticle array with nanoparticle diameters of 300 and 250 nm can be distinguished.In addition,the Talbot effect of the 300-nm-diameter nanoparticle array is also observed.If a nanoparticle sample is assembled on a glass slide deposited with a 30-nm-thick Ag film,an array of 300-nm-diameter nanoparticles can be discerned.We propose that in microsphere-assisted microscopy imaging,the resolution can be improved by the excitation of surface plasmon polaritons(SPPs) on the sample surface or at the sample/substrate interface,and a higher near-field intensity due to the excited SPPs would benefit the resolution improvement.Our study of label-free super-resolution imaging of low-contrast objects will promote the applications of microsphere-assisted microscopy in life sciences.展开更多
Microvascular structure and hemodynamics are important indicators for the diagnosis and assessment of many diseases and pathologies.The structural and functional imaging of tissue microvasculature in vivo is a clinica...Microvascular structure and hemodynamics are important indicators for the diagnosis and assessment of many diseases and pathologies.The structural and functional imaging of tissue microvasculature in vivo is a clinically significant objective for the development of many imaging modalities.Contrast-enhanced ultrasound(CEUS)is a popular clinical tool for characterizing tissue microvasculature,due to the moderate cost,wide accessibility,and absence of ionizing radiation of ultrasound.展开更多
Axial excitation confinement beyond the diffraction limit is crucial to the development of next-generation,super-resolution microscopy.STimulated Emission Depletion(STED)nanoscopy offers lateral super-resolution using...Axial excitation confinement beyond the diffraction limit is crucial to the development of next-generation,super-resolution microscopy.STimulated Emission Depletion(STED)nanoscopy offers lateral super-resolution using a donut-beam depletion,but its axial resolution is still over 500 nm.Total internal reflection fluorescence microscopy is widely used for single-molecule localization,but its ability to detect molecules is limited to within the evanescent field of~100 nm from the cell attachment surface.We find here that the axial thickness of the point spread function(PSF)during confocal excitation can be easily improved to 110 nm by replacing the microscopy slide with a mirror.The interference of the local electromagnetic field confined the confocal PSF to a 110-nm spot axially,which enables axial super-resolution with all laser-scanning microscopes.Axial sectioning can be obtained with wavelength modulation or by controlling the spacer between the mirror and the specimen.With no additional complexity,the mirror-assisted excitation confinement enhanced the axial resolution six-fold and the lateral resolution two-fold for STED,which together achieved 19-nm resolution to resolve the inner rim of a nuclear pore complex and to discriminate the contents of 120 nm viral filaments.The ability to increase the lateral resolution and decrease the thickness of an axial section using mirror-enhanced STED without increasing the laser power is of great importance for imaging biological specimens,which cannot tolerate high laser power.展开更多
基金supported by the National Natural Science Foundation of China(62175034,62175036,32271510)the National Key R&D Program of China(2021YFF0502900)+2 种基金the Science and Technology Research Program of Shanghai(Grant No.19DZ2282100)the Shanghai Key Laboratory of Metasurfaces for Light Manipulation(23dz2260100)the Shanghai Engineering Technology Research Center of Hair Medicine(19DZ2250500).
文摘A new scheme of super-resolution optical fluctuation imaging(SOFI)is proposed to broaden its application in the high-order cumulant reconstruction by optimizing blinking characteristics,eliminating noise in raw data and applying multi-resolution analysis in cumulant reconstruction.A motor-driven rotating mask optical modulation system is designed to adjust the excitation lightfield and allows for fast deployment.Active-modulated fluorescence fluctuation superresolution microscopy with multi-resolution analysis(AMF-MRA-SOFI)demonstrates enhanced resolution ability and reconstruction quality in experiments performed on sample of conventional dyes,achieving a resolution of 100 nm in the fourth order compared to conventional SOFI reconstruction.Furthermore,our approach combining expansion super-resolution achieved a resolution at-57 nm.
基金supported by the following grants:National Key R&D Program of China(Grant no.2022YFC3401100)National Natural Science Foundation of China(Grant nos.32271428,92054110,32201132 and 31600692).
文摘Blood cells are the most integral part of the body,which are made up of erythrocytes,platelets and white blood cells.The examination of subcellular structures and proteins within blood cells at the nanoscale can provide valuable insights into the health status of an individual,accurate diagnosis,and efficient treatment strategies for diseases.Super-resolution microscopy(SRM)has recently emerged as a cutting-edge tool for the study of blood cells,providing numerous advantages over traditional methods for examining subcellular structures and proteins.In this paper,we focus on outlining the fundamental principles of various SRM techniques and their applications in both normal and diseased states of blood cells.Furthermore,future prospects of SRM techniques in the analysis of blood cells are also discussed.
基金support from National Basic Research Program of China (973 Program) (2015CB352005)National Natural Science Foundation of China (No.6142780065,31571110,81527901)+1 种基金Natural Science Foundation of Zhejiang Province of China (No.Y16F050002)the Fundamental Research Funds for the Central Universities.
文摘Optical microscopy promises researchers to soe most tiny substances directly.However,the resolution of conventional microscopy is resticted by the diffraction limit.This makes it a challenge to observe subcellular processes happened in nanoscale.The development of super-resolution microscopy provides a solution to this challenge.Here,we briefly review several commonly used super-resolution techniques,explicating their basic principles and applications in biological science,especially in neuroscience.In addition,characteristics and limitations of each techrique are compared to provide a guidance for biologists to choose the most suitable tool.
基金sponsored by National Natural Science Foundation of China(61827825 and 61735017)Fundamental Research Funds for the Central Universities(2019XZZX003-06)+1 种基金Natural Science Foundation of Zhejiang province(LR16F050001)Zhejiang Lab(2018EB0ZX01).
文摘Image scanning microscopy based on pixel reassignment can improve the confocal resolution limit without losing the image signal-to-noise ratio(SNR)greatly[C.J.R.Sheppard,"Super resolution in confocal imaging,"Optik 80(2)53-54(1988).C.B.Miller,E.Jorg,"Image scanning microscopy,"Phys.Reu.Lett.104(19)198101(2010).C.J.R.Sheppard,s.B.Mehta,R Heintzmann,"Superresolution by image scanning microscopy using pixel reassignment,"Opt.Lett.38(15)28892892(2013)].Here,we use a tailor-made optical fiber and 19 avalanche pho-todiodes(APDs)as parallel detectors to upgrade our existing confocal microscopy,termed as parallel-detection super resolution(PDSR)microscopy.In order to obtain the correct shift value,we use the normalized 2D cross correlation to calculate the shifting value of each image.We characterized our system performance by imaging fuorescence beads and applied this system to observing the 3D structure of biological specimen.
基金This study was supported by:Inner Mongolia Academy of Forestry Sciences Open Research Project(Grant No.KF2024MS03)The Project to Improve the Scientific Research Capacity of the Inner Mongolia Academy of Forestry Sciences(Grant No.2024NLTS04)The Innovation and Entrepreneurship Training Program for Undergraduates of Beijing Forestry University(Grant No.X202410022268).
文摘Remote sensing image super-resolution technology is pivotal for enhancing image quality in critical applications including environmental monitoring,urban planning,and disaster assessment.However,traditional methods exhibit deficiencies in detail recovery and noise suppression,particularly when processing complex landscapes(e.g.,forests,farmlands),leading to artifacts and spectral distortions that limit practical utility.To address this,we propose an enhanced Super-Resolution Generative Adversarial Network(SRGAN)framework featuring three key innovations:(1)Replacement of L1/L2 loss with a robust Charbonnier loss to suppress noise while preserving edge details via adaptive gradient balancing;(2)A multi-loss joint optimization strategy dynamically weighting Charbonnier loss(β=0.5),Visual Geometry Group(VGG)perceptual loss(α=1),and adversarial loss(γ=0.1)to synergize pixel-level accuracy and perceptual quality;(3)A multi-scale residual network(MSRN)capturing cross-scale texture features(e.g.,forest canopies,mountain contours).Validated on Sentinel-2(10 m)and SPOT-6/7(2.5 m)datasets covering 904 km2 in Motuo County,Xizang,our method outperforms the SRGAN baseline(SR4RS)with Peak Signal-to-Noise Ratio(PSNR)gains of 0.29 dB and Structural Similarity Index(SSIM)improvements of 3.08%on forest imagery.Visual comparisons confirm enhanced texture continuity despite marginal Learned Perceptual Image Patch Similarity(LPIPS)increases.The method significantly improves noise robustness and edge retention in complex geomorphology,demonstrating 18%faster response in forest fire early warning and providing high-resolution support for agricultural/urban monitoring.Future work will integrate spectral constraints and lightweight architectures.
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 Ministry of Science,ICT and Future Planning(MSIP)through the National Research Foundation of Korea(NRF)(RS-2024-00450201)supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Ministry of Health and Welfare,Republic of Korea(HI22C1374).
文摘We developed an imaging technique combining two-photon computed super-resolution microscopy and suction-based stabilization to achieve the resolution of the single-cell level and organelles in vivo.To accomplish this,a conventional two-photon microscope was equipped with a 3D-printed holders,which stabilize the tissue surface within the focal plane of immersion objectives.Further computational image stabilization and noise reduction were applied,followed by superresolution radial fluctuations(SRRF)analysis,doubling image resolution,and enhancing signal-to-noise ratios for in vivo subcellular process investigation.Stabilization of<1μm was obtained by suction,and<25 nm were achieved by subsequent algorithmic image stabilization.A Mito-Dendra2 mouse model,expressing green fluorescent protein(GFP)in mitochondria,demonstrated the potential of long-term intravital subcellular imaging.In vivo mitochondrial fission and fusion,mitochondrial status migration,and the effects of alcohol consumption(modeled as an alcoholic liver disease)and berberine treatment on hepatocyte mitochondrial dynamics are directly observed intravitally.Suction-based stabilization in two-photon intravital imaging,coupled with computational super-resolution holds promise for advancing in vivo subcellular imaging studies.
基金Project supported by the National Key Foundation for Exploring Scientific Instrument （No. 2013YQ03065102）, the National Basic Research Program （973） of China （No. 2012CB316503）, and the National Natural Science Foundation of China （Nos. 31327901, 61475010, 31361163004, and 61428501）
文摘The broad applicability of super-resolution microscopy has been widely demonstrated in various areas and disciplines. The optimization and improvement of algorithms used in super-resolution microscopy are of great importance for achieving optimal quality of super-resolution imaging. In this review, we comprehensively discuss the computational methods in different types of super-resolution microscopy, including deconvolution microscopy, polarization-based super-resolution microscopy, structured illumination microscopy, image scanning microscopy, super-resolution optical fluctuation imaging microscopy, single-molecule localization microscopy, Bayesian super-resolution microscopy, stimulated emission depletion microscopy, and translation microscopy. The development of novel computational methods would greatly benefit super-resolution microscopy and lead to better resolution, improved accuracy, and faster image processing.
基金supported by the National Natural Science Foundation of China (81427801, 81827901)National Basic Research Program of China (2015CB352003)+2 种基金Science Fund for Creative Research Groups (61721092)Fundamental Research Funds for the Central Universities (2018KFYXKJC039)Director Fund of WNLO。
文摘Resolution is undoubtedly the most important parameter in optical microscopy by providing an estimation on the maximum resolving power of a certain optical microscope. For centuries, the resolution of an optical microscope is generally considered to be limited only by the numerical aperture of the optical system and the wavelength of light. However, since the invention and popularity of various advanced fluorescence microscopy techniques, especially super-resolution fluorescence microscopy, many new methods have been proposed for estimating the resolution, leading to confusions for researchers who need to quantify the resolution of their fluorescence microscopes. In this paper, we firstly summarize the early concepts and criteria for predicting the resolution limit of an ideal optical system. Then, we discuss some important influence factors that deteriorate the resolution of a certain fluorescence microscope. Finally, we provide methods and examples on how to measure the resolution of a fluorescence microscope from captured fluorescence images. This paper aims to answer as best as possible the theoretical and practical issues regarding the resolution estimation in fluorescence microscopy.
基金the National Natural Science Foundation of China(91850202,92150301,12074121,62105101,62175066,11727810,12034008,12274129,12274139)Science and Technology Commission of Shanghai Municipality(21XD1400900,20ZR1417100,21JM0010700).
文摘Various super-resolution microscopy techniques have been presented to explore fine structures of biological specimens.However,the super-resolution capability is often achieved at the expense of reducing imaging speed by either point scanning or multiframe computation.The contradiction between spatial resolution and imaging speed seriously hampers the observation of high-speed dynamics of fine structures.To overcome this contradiction,here we propose and demonstrate a temporal compressive super-resolution microscopy(TCSRM)technique.This technique is to merge an enhanced temporal compressive microscopy and a deep-learning-based super-resolution image reconstruction,where the enhanced temporal compressive microscopy is utilized to improve the imaging speed,and the deep-learning-based super-resolution image reconstruction is used to realize the resolution enhancement.The high-speed super-resolution imaging ability of TCSRM with a frame rate of 1200 frames per second(fps)and spatial resolution of 100 nm is experimentally demonstrated by capturing the flowing fluorescent beads in microfluidic chip.Given the outstanding imaging performance with high-speed super-resolution,TCSRM provides a desired tool for the studies of high-speed dynamical behaviors in fine structures,especially in the biomedical field.
基金supported by the National Natural Science Foundation of China(Grant Nos.62122028 and 62335008)Guangdong Basic and Applied Basic Research Foundation(Grant Nos.2023B1515040018 and 2018B030306015)+1 种基金National Key Research and Development Program of China(Grant No.2023YFF0722600)Scientific Research Cultivation Fund for Young Faculty of South China Normal University(Grant Nos.23KJ01 and 22KJ30).
文摘Photon avalanche occurring in lanthanide-doped materials exhibits a giant optical nonlinear response of the emission intensity to the excitation intensity,which holds great potential in the applications of optical sensing,super-resolution imaging,quantum detection,and other techniques.However,strategies for developing photon avalanches in nanoparticles are limited,and many widely used lanthanide ions have not yet been able to generate high-efficiency avalanching emissions.A general strategy named cascade migrating photon avalanche was proposed to achieve efficient avalanching emissions with huge optical nonlinearities from a large number of emitters at the nanoscale and at room temperature.Specifically,the optical nonlinearity order of bright avalanched Tm^(3+)-emission was achieved at 63rd order by utilizing the Yb^(3+)∕Pr^(3+)-codoped nano-engine.By further incorporating a Gd^(3+)sublattice migrating network,its avalanching energy can propagate over a long distance to arouse avalanching emission with extreme optical nonlinearities up to 45th order among various emitters(Tb^(3+),Eu^(3+),Dy^(3+),Sm^(3+))in multilayered nanostructures.By achieving abundant avalanching full-spectrum emissions,it would be highly conducive to applications in various fields.For instance,our strategy demonstrated its applicability in multi-color super-resolution microscopic imaging with single-nanoparticle sensitivity and resolution up to 48 nm,utilizing a single low-power 852 nm excitation beam.
基金supported by the National Key Research and Development Program of China(2022YFC3400600)the National Natural Science Foundation of China(11874231,32227802,and 12174208)+3 种基金Guangdong Major Project of Basic and Applied Basic Research(2020B0301030009)China Postdoctoral Science Foundation(2020M680032)the Fundamental Research Funds for the Central Universities(2122021337 and 2122021405)the Pew Charitable Trusts.
文摘The transmembrane protein CD47,an innate immune checkpoint protein,plays a pivotal role in preventing healthy erythrocytes from immune clearance.Our study utilized stochastic optical reconstruction microscopy(STORM)and single-molecule analysis to investigate the distribution of CD47 on the human erythrocyte membrane.Contrary to previous findings in mouse erythrocytes,we discovered that CD47 exists in randomly distributed monomers rather than in clusters across the human erythrocyte membrane.Using secondary antibody-induced crosslinking,we found that CD47 aggregates into stable clusters within minutes.By comparing these STORM results with those of the fully mobile protein CD59 and the cytoskeleton-bound membrane protein glycophorin C under similar conditions,as well as devising two-color STORM co-labeling and co-clustering experiments,we further quantitatively revealed an intermediate,self-limiting clustering behavior of CD47,elucidating its fractional(∼14%)attachment to the cytoskele-ton.Moreover,we report reductions in both the amount of CD47 and its clustering capability in aged erythrocytes,providing new insight into erythrocyte senescence.Together,the combination of STORM and secondary antibody-based crosslinking unveils the unique self-limiting clustering behavior of CD47 due to its fractional cytoskeleton attachment.
文摘With the support by the National Natural Science Foundation of China and the Chinese Academy of Sciences,the research team led by Prof.Li Junbai(李峻柏)at the CAS Key Lab of Colloid,Interface and Thermodynamics,Institute of Chemistry,Chinese Academy of Sciences,revealed the distribution of proteins in the transformation of inorganic/protein hybrid crystals by super-resolution microscopy,which
基金support from the National Institutes of Health(Grant R35GM133505)the National Science Foundation(Grant no.2237129)the University of Houston.
文摘The introduction of super-resolution microscopy(SRM)has significantly advanced our understanding of cellular and molecular dynamics,offering a detailed view previously beyond our reach.Implementing SRM in biophysical research,however,presents numerous challenges.This review addresses the crucial aspects of utilizing SRM effectively,from selecting appropriate fluorophores and preparing samples to analyzing complex data sets.We explore recent technological advancements and methodological improvements that enhance the capabilities of SRM.Emphasizing the integration of SRM with other analytical methods,we aim to overcome inherent limitations and expand the scope of biological insights achievable.By providing a comprehensive guide for choosing the most suitable SRM methods based on specific research objectives,we aim to empower researchers to explore complex biological processes with enhanced precision and clarity,thereby advancing the frontiers of biophysical research.
基金supported by the National Key R&D Program of China(2021YFF0502900)the National Natural Science Foundation of China(61835009/62127819).
文摘The algorithm used for reconstruction or resolution enhancement is one of the factors affectingthe quality of super-resolution images obtained by fluorescence microscopy.Deep-learning-basedalgorithms have achieved stateof-the-art performance in super-resolution fluorescence micros-copy and are becoming increasingly attractive.We firstly introduce commonly-used deep learningmodels,and then review the latest applications in terms of the net work architectures,the trainingdata and the loss functions.Additionally,we discuss the challenges and limits when using deeplearning to analyze the fluorescence microscopic data,and suggest ways to improve the reliability and robustness of deep learning applications.
文摘The resolution of conventional optical microscopy is only -200 nm, which is becoming less and less sufficient for a variety of applications. In order to surpass the diffraction limited resolution, super-resolution microscopy （SRM） has been developed to achieve a high resolution of one to tens of nanometers. The techniques involved in SRM can be assigned into two broad categories, namely ＂true＂ super-resolution techniques and ＂functional＂ super-resolution techniques. In ＂functional＂ super-resolution techniques, stochastic super-resolution microscopy （SSRM） is widely used due to its low expense, simple operation, and high resolution. The principle process in SSRM is to accumulate the coordinates of many diffraction-limited emitters （e.g., single fluorescent molecules） on the object by localizing the centroids of the point spread functions （PSF）, and then reconstruct the image of the object using these coordinates. When the diffraction-limited emitters take part in a catalytic reaction, the activity distribution and kinetic information about the catalysis by nanoparticles can be obtained by SSRM. SSRM has been applied and exhibited outstanding advantages in several fields of catalysis, such as metal nanoparticle catalysis, molecular sieve catalysis, and photocatalysis. Since SSRM is able to resolve the catalytic activity within one nanoparticle, it promises to accelerate the development and discovery of new and better catalysts. This review will present a brief introduction to SRM, and a detailed description of SSRM and its applications in nano-catalysis.
基金This work was supported by the Innovation Fund of WNLO(2018WNLOKF023)the Start-up Fund of Hainan University(KYQD(ZR)-20077).
文摘Hematologic malignancies are one of the most common malignant tumors caused by the clonal proliferation and differentiation of hematopoietic and lymphoid stem cells.The examination of bone marrow cells combined with immunodeficiency typing is of great significance to the diagnostic type,treatment and prognosis of hematologic malignancies.Super-resolution fluorescence microscopy(SRM)is a special kind of optical microscopy technology,which breaks the resolution limit and was awarded the Nobel Prize in Chemistry in 2014.With the development of SRM,many related technologies have been applied to the diagnosis and treatment of clinical diseases.It was reported that a major type of SRM technique,single molecule localization microscopy(SMLM),is more sensitive than flow cytometry(FC)in detecting cell membrane antigens'expression,thus enabling better chances in detecting antigens on hematopoietic cells than traditional analytic tools.Furthermore,SRM may be applied to clinical pathology and may guide precision medicine and personalized medicine for clone hematopoietic cell diseases.In this paper,we mainly discuss the application of SRM in clone hematological malignancies.
基金Project supported by the National Natural Science Foundation of China(Grant No.61673287)。
文摘We use the label-free microsphere-assisted microscopy to image low-contrast hexagonally close-packed polystyrene nanoparticle arrays with diameters of 300 and 250 nm.When a nanoparticle array is directly placed on a glass slide,it cannot be distinguished.If a 30-nm-thick Ag film is deposited on the surface of a nanoparticle array,the nanoparticle array with nanoparticle diameters of 300 and 250 nm can be distinguished.In addition,the Talbot effect of the 300-nm-diameter nanoparticle array is also observed.If a nanoparticle sample is assembled on a glass slide deposited with a 30-nm-thick Ag film,an array of 300-nm-diameter nanoparticles can be discerned.We propose that in microsphere-assisted microscopy imaging,the resolution can be improved by the excitation of surface plasmon polaritons(SPPs) on the sample surface or at the sample/substrate interface,and a higher near-field intensity due to the excited SPPs would benefit the resolution improvement.Our study of label-free super-resolution imaging of low-contrast objects will promote the applications of microsphere-assisted microscopy in life sciences.
基金This study was supported by the National Cancer Institute(NCI)of the National Institutes of Health(NIH)under Award(No.R00CA214523).
文摘Microvascular structure and hemodynamics are important indicators for the diagnosis and assessment of many diseases and pathologies.The structural and functional imaging of tissue microvasculature in vivo is a clinically significant objective for the development of many imaging modalities.Contrast-enhanced ultrasound(CEUS)is a popular clinical tool for characterizing tissue microvasculature,due to the moderate cost,wide accessibility,and absence of ionizing radiation of ultrasound.
基金supported by the National Instrument Development Special Program(2013YQ03065102)the‘973’Major State Basic Research Development Program of China(2011CB809101)+2 种基金the Natural Science Foundation of China(31327901,61475010,61428501)the Australian Research Council Centre of Excellence for Nanoscale BioPhotonics(CE140100003)provided by the National Institute of Health(GM094198 to PJS).
文摘Axial excitation confinement beyond the diffraction limit is crucial to the development of next-generation,super-resolution microscopy.STimulated Emission Depletion(STED)nanoscopy offers lateral super-resolution using a donut-beam depletion,but its axial resolution is still over 500 nm.Total internal reflection fluorescence microscopy is widely used for single-molecule localization,but its ability to detect molecules is limited to within the evanescent field of~100 nm from the cell attachment surface.We find here that the axial thickness of the point spread function(PSF)during confocal excitation can be easily improved to 110 nm by replacing the microscopy slide with a mirror.The interference of the local electromagnetic field confined the confocal PSF to a 110-nm spot axially,which enables axial super-resolution with all laser-scanning microscopes.Axial sectioning can be obtained with wavelength modulation or by controlling the spacer between the mirror and the specimen.With no additional complexity,the mirror-assisted excitation confinement enhanced the axial resolution six-fold and the lateral resolution two-fold for STED,which together achieved 19-nm resolution to resolve the inner rim of a nuclear pore complex and to discriminate the contents of 120 nm viral filaments.The ability to increase the lateral resolution and decrease the thickness of an axial section using mirror-enhanced STED without increasing the laser power is of great importance for imaging biological specimens,which cannot tolerate high laser power.
