Static recording characteristics of new type super-resolution near-field structure

A novel super-resolution near-field optical structure （super-RENS） with bismuth （Bi） mask layer is proposed in this paper. Static optical recording tests with and without super-RENS are carried out using a 650-nm semiconductor laser at recording powers of 14 and 7 mW with pulse duration of 100 ns. The recording marks are observed by high-resolution optical microscopy with a charge-coupled device （CCD） camera. The results show that the Bi mask layer can also concentrate energy into the center of a laser beam at low laser power similar to the traditional Sb mask layer. The results above are further confirmed by another Ar;laser system. The third-order nonlinear response induced by the plasma oscillation at the Bi/SiN interface during laser irradiation can be used to explain the phenomenon. The calculation results are basically consistent with our experimental results.

Progress of super-resolution near-field structure and application in optical data storage

The era of big data has necessitated the use of ultra-high density optical storage devices. Super-resolution near-field structure （super-RENS）, which has successfully surpassed the fundamental optical diffraction limit, is one of the promising next generation high-density optical storage technologies. This technology combines the traditional super-resolution optical disk with a near-field structure, and has the advantages of structural simplicity, strong practicability, and, more importantly, compatibility with the current optical storage pickup. The mask layer in super-RENS functions as the key to realizing superresolution. Development of suitable materials and stack designs has greatly been improved in the last decade. This paper described several types of super-RENS, such as aperture-type, light scattering center-type, bubble-type, and other types （e.g., WOx and ZnO）, particularly the newly proposed super-RENS technology and research achievements. The paper also reviews the applications of super-RENS in high-density optical data storage in recent years. After analyzing and comparing various types of super-RENS technology, the paper proposes the aperturetype based on the mechanism of nonlinear optics as the most suitable candidate for practical applications in the near future.

Fringe optimization for structured illumination super-resolution microscope with digital micromirror device

Structured illumination microscopy(SIM)is a promising super-resolution technique for imaging subcellular structures and dynamics due to its compatibility with most commonly usedffuorescent labeling methods.Structured illumination can be obtained by either laser interference or projection of fringe patterns.Here,we proposed a fringe projector composed of a compact multiwavelength LEDs module and a digital micromirror device(DMD)which can be directly attached to most commercial invertedffuorescent microscopes and update it into a SIM system.The effects of the period and duty cycle of fringe patterns on the modulation depth of the structured lightfield were studied.With the optimized fringe pattern,1:6×resolution improvement could be obtained with high-end oil objectives.Multicolor imaging and dynamics of subcellular organelles in live cells were also demonstrated.Our method provides a low-cost solution for SIM setup to expand its wide range of applications to most research labs in thefield of life science and medicine.

Design of Network Cascade Structure for Image Super-Resolution

Image super resolution is an important field of computer research.The current mainstream image super-resolution technology is to use deep learning to mine the deeper features of the image,and then use it for image restoration.However,most of these models mentioned above only trained the images in a specific scale and do not consider the relationships between different scales of images.In order to utilize the information of images at different scales,we design a cascade network structure and cascaded super-resolution convolutional neural networks.This network contains three cascaded FSRCNNs.Due to each sub FSRCNN can process a specific scale image,our network can simultaneously exploit three scale images,and can also use the information of three different scales of images.Experiments on multiple datasets confirmed that the proposed network can achieve better performance for image SR.

A lateral super-resolution imaging method using structured illumination without phase shift

Structured illumination microscopy has been a useful method for achieving lateral super-resolution,but it typically requires at least three precise phase shifts per orientation.In this paper,we propose a super-resolution method that utilizes structured illumination without phase shift.The reconstruction process requires only a conventionally illuminated image and an image with structured illumination.This method achieves the same effect as the traditional phase shift method,and more than doubles the resolution by synthesizing a few reconstructions at different illumination frequencies.We verify the resolution improvement process using a combination of theoretical derivations and diagrams,and demonstrate its effectiveness with numerical simulations.

Comparison of point detection and area detection for point-scanning structured illumination microscopy

Structured illumination microscopy(SIM)is suitable for biological samples because of its relatively low-peak illumination intensity requirement and high imaging speed.The system resolution is affected by two typical detection modes:Point detection and area detection.However,a systematic analysis of the imaging performance of the different detection modes of the system has rarely been conducted.In this study,we compared laser point scanning point detection(PS-PD)and point scanning area detection(PS-AD)imaging in nonconfocal microscopy through theoretical analysis and simulated imaging.The results revealed that the imaging resolutions of PSPD and PS-AD depend on excitation and emission point spread functions(PSFs),respectively.Especially,we combined the second harmonic generation(SHG)of point detection(P-SHG)and area detection(A-SHG)with SIM to realize a nonlinear SIM-imaging technique that improves the imaging resolution.Moreover,we analytically and experimentally compared the nonlinear SIM performance of P-SHG with that of A-SHG.

Nonlinear scanning structured illumination microscopy based on nonsinusoidal modulation

Structured illumination microscopy(SIM)is an essential super-resolution microscopy technique that enhances resolution.Several images are required to reconstruct a super-resolution image.However,linear SIM resolution enhancement can only increase the spatial resolution of micros-copy by a factor of two at most because the frequency of the structured illumination pattern is limited by the cutoff frequency of the excitation point spread function.The frequency of the pattern generated by the nonlinear response in samples is not limited;therefore,nonlinear SIM(NL-SIM),in theory,has no inherent limit to the resolution.In the present study,we describe a two-photon nonlinear SIM(2P-SIM)technique using a multiple harmonics scanning pattern that employs a composite structured illumination pattern,which can produce a higher order harmonic pattern based on the fluorescence nonlinear response in a 2P process.The theoretical models of super-resolution imaging were established through our simulation,which describes the working mechanism of the multi-frequency structure of the nonsinusoidal function to improve the reso-lution.The simulation results predict that a 5-fold improvement in resolution in the 2P-SIM is possible.

Structured illumination microscopy based on asymmetric three-beam interference

Structured illumination microscopy(SIM)is a rapidly developing super-resolution technology.It has been widely used in various application fields of biomedicine due to its excellent two-and three-dimensional imaging capabilities.Furthermore,faster three-dimensional imaging methods are required to help enable more research-oriented living cell imaging.In this paper,a fast and sensitive three-dimensional structured illumination microscopy based on asymmetric three-beam interference is proposed.An innovative time-series acquisition method is employed to halve the time required to obtain each raw image.A segmented half-wave plate as a substantial linear polarization modulation method is applied to the three-dimensional SIM system for the first time.Although it needs to acquire 21 raw images instead of 15 to reconstruct one super-resolution image,the SIM setup proposed in this paper is 30%faster than the traditional spatial light modulator-SIM(SLM-SIM)in imaging each super-resolution image.The related theoretical derivation,hardware system,and verification experiment are elaborated in this paper.The stable and fast 3D super-resolution imaging method proposed in this paper is of great significance to the research of organelle interaction,intercellular communication,and other biomedical fields.

Structured illumination microscopy and its new developments

Optical microscopy allows us to observe the biological structures and processes within living cells.However,the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light di®raction.Structured illumination microscopy(SIM),a type of new emerging super-resolution microscopy,doubles the spatial resolution by illuminating the specimen with a patterned light,and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy.In addition,SIM is easier to combine with the other imaging techniques to improve their imaging resolution,leading to the developments of diverse types of SIM.SIM has great potential to meet the various requirements of living cells imaging.Here,we review the recent developments of SIM and its combination with other imaging techniques.

Improvement in Resolution of Multiphoton Scanning Structured Illumination Microscopy via Harmonics

We describe a multiphoton(mP)-structured illumination microscopy(SIM)technique,which demonstrates substantial improvement in image resolution compared with linear SIM due to the nonlinear response of fluorescence.This nonlinear response is caused by the effect of nonsinusoidal structured illumination created by scanning a sinusoidally modulated illumination to excite an mP fluorescence signal.The harmonics of the structured fluorescence illumination are utilised to improve resolution.We present an mP-SIM theory for reconstructing the super-resolution image of the system.Theoretically,the resolution of our m P-SIM is unlimited if all the high-order harmonics of the nonlinear response of fluorescence are considered.Experimentally,we demonstrate an 86 nm lateral resolution for two-photon(2P)-SIM and a 72 nm lateral resolution for second-harmonic-generation(SHG)-SIM.We further demonstrate their application by imaging cells stained with F-actin and collagen fibres in mouse-tail tendon.Our method can be directly used in commercial mP microscopes and requires no specific fluorophores or high-intensity laser.

Application of super-resolution fluorescence microscopy in hematologic malignancies

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.

Super-Resolution Image Reconstruction Based on an Improved Maximum a Posteriori Algorithm

A maximum a posteriori（ MAP） algorithm is proposed to improve the accuracy of super resolution（ SR） reconstruction in traditional methods. The algorithm applies both joints image registration and SR reconstruction in the framework,but separates them in the process of iteratiion. Firstly,we estimate the shifting parameters through two lowresolution（ LR） images and use the parameters to reconstruct initial HR images. Then,we update the shifting parameters using HR images. The aforementioned steps are repeated until the ideal HR images are obtained. The metrics such as PSNR and SSIM are used to fully evaluate the quality of the reconstructed image. Experimental results indicate that the proposed method can enhance image resolution efficiently.

Method of lateral image reconstruction in structured illumination microscopy with super resolution

The image reconstruction process in super-resolution structured illumination microscopy(SIM)is investigated.The structured pattern is generated by the interference of two Gaussian beams to encode undetectable spectra into detectable region of microscope.After parameters estimation of the structured pattern,the encoded spectra are computationally decoded and recombined in Fourier domain to equivalently increase the cut-off frequency of microscope,resulting in the extension of detectable spectra and a reconstructed image with about two-fold enhanced resolution.Three di®erent methods to estimate the initial phase of structured pattern are compared,verifying the auto-correlation algorithm a®ords the fast,most precise and robust measurement.The artifacts sources and detailed reconstruction°owchart for both linear and nonlinear SIM are also presented.

Super-resolution microscopy and its applications in neuroscience

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.

Progress and trend on near-field problems in civil engineering

In the last twenty years, near-field problems became an important topic for both seismologists and civil engineers. The one aspect is to illuminate mechanisms of earthquakes and explain new phenomena. The another aspect is the ground motions, which are usually assigned by engineers as a type of input load for seismic design of structures, sometimes can control the final design results. The experiments, performance evaluations and other related aspects are all based on the specified type of load. As a result, many aspects related to civil engineering will be influenced by changes of the type of load, Hence, the characteristics of the load and the corresponding response of structures are desired for studying. In this paper, the state-of-the-art of near-field problems in civil engineering is comprehensively reviewed, which include inherent characteristics of near-field ground motions and influences of these ground motions on civil structures. The existing problems are pointed out and work needed to be further investigated in the future is suggested. It is believed that the information in this paper can be useful to advance the state of investigation on near-field problems.

Enhancement of Near-Field Radiative Heat Transfer based on High-Entropy Alloys

The enhancement of near-field radiative heat transfer(NFRHT)has now become one of the research hotspots in the fieldsof thermal management and imaging due to its ability to improve the performance of near-field thermoelectric devices and near-field imaging systems.In this paper,we design three structures(multilayer structure,nanoporous structure,and nanorod structure)based on high-entropy alloys to realize the enhancement of NFRHT.By combining stochastic electrodynamicsand Maxwell-Garnett's description of the effective medium,we calculate the radiative heat transfer under different parametersand find that the nanoporousstructure has the largest enhancement effect on NFRHT.The near-field heat transfer factor(q)of this structure(q=1.40×10^(9)W/(m^(2)·K))is three times higher than that of the planestructure(q=4.6×10^(8)W/(m^(2)·K)),and about two orders of magnitude higher than that of the SiO2plate.Thisresult providesa freshidea for the enhancement of NFRHT and will promote the application of high-entropy alloy materials in near-field heat radiation.

Fluorescence interference structured illumination microscopy for 3D morphology imaging with high axial resolution

Imaging three-dimensional,subcellular structures with high axial resolution has always been the core purpose of fluorescence microscopy.However,trade-offs exist between axial resolution and other important technical indicators,such as temporal resolution,optical power density,and imaging process complexity.We report a new imaging modality,fluorescence interference structured illumination microscopy(FI-SIM),which is based on three-dimensional structured illumination microscopy for wide-field lateral imaging and fluorescence interference for axial reconstruction.FI-SIM can acquire images quickly within the order of hundreds of milliseconds and exhibit even 30 nm axial resolution in half the wavelength depth range without z-axis scanning.Moreover,the relatively low laser power density relaxes the requirements for dyes and enables a wide range of applications for observing fixed and live subcellular structures.

Super-resolution imaging reveals the subcellular distribution of dextran at the nanoscale in living cells

Theranostic visualization of dextran at the nanoscale is beneficial for understanding the bioregulatory mechanisms of this molecule. In this study, we applied structured illumination microscopy(SIM) to capture the distribution of Cy5-Dextran at different incubation periods in living cells. The results showed that Cy5-Dextran could be absorbed by He La cells. In addition, we clarified that Cy5-Dextran exhibited differential organelle distribution(lysosomal or mitochondrial) in a time-dependent manner. Moreover,lysosomal Cy5-Dextran localization was found to be independent of the autophagy process, while Cy5-Dextran localized to the mitochondria triggered a pro-apoptotic event, upregulating the levels of reactive oxygen species(ROS) to accelerate mitochondrial fragmentation. This work uses a visualized strategy to reveal the anti-tumor bioactivity of dextran, which was achieved by regulating apoptosis and autophagy.

Donor-Acceptor Typed AIE Luminogens with Near-infrared Emission for Super-resolution Imaging

Aggregation-induced emission(AIE)luminogens(AIEgens)with high brightness in aggregates exhibit great potentials in biological imaging,but these AIEgens are seldom applied in super-resolution biological imaging,especially in the imaging by using the structural illumination microscope(SIM).Based on this consideration,we synthesized the donor-acceptor typed AIEgen of DTPA-BTN,which not only owns high brightness in the near-infrared(NIR)emission region from 600 nm to 1000 nm(photoluminescence quantum yield,PLQYs=11.35%),but also displays excellent photo-stability.In addition,AIE nanoparticles based on 4,7-ditriphenylamine-[1,2,5]-thiadiazolo[3,4-c]pyridine(DTPA-BTN)were also prepared with highly emissive features and excellent biocompatibility.Finally,the developed DTPA-BTN-based AIE nanoparticles were applied in the super-resolution cellular imaging via SIM,where much smaller full width at half-maximum values and high signal to noise ratios were obtained,indicating the superior imaging resolution.The results here imply that highly emissive AIEgens or AIE nanoparticles can be promising imaging agents for super-resolution imaging via SIM.

Illuminating the structure and dynamics of chromatin by fluorescence labeling

BACKGROUND： Visualization of chromosomal loci location and dynamics is crucial for understanding many fundamental intra-nuclear processes such as DNA transcription, replication, and repair. OBJECTIVE： Here, we will describe the development of fluorescence labeling methods for chromatin imaging, including traditional as well as emerging chromatin labeling techniques in both fixed and live cells. We will also discuss current issues and provide a perspective on future developments and applications of the chromatin labeling technology. METHODS： A systematic literature search was performed using the PubMed. Studies published over the past 50 years were considered for review. More than 100 articles were cited in this review. RESULTS： Taking into account sensitivity, specificity, and spatiotemporal resolution, fluorescence labeling and imaging has been the most prevalent approach for chromatin visualization. Among all the fluorescent labeling tools, the adoption ofgenome editing tools, such as TALE and CRISPR, have great potential for the labeling and imaging of chromatin. CONCLUSION： Although a number of chromatin labeling techniques are available for both fixed and live cells, much more effort is still clearly required to develop fluorescence labeling methods capable of targeting arbitrary sequences non-intrusively to allow long-term, multiplexing, and high-throughput imaging of genomic loci and chromatin structures. The emerging technological advances will outline a next-generation effort toward the comprehensive delineation of chromatin at single-cell level with single-molecule resolution.