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Estimation-free spatial-domain image reconstruction of structured illumination microscopy

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Journal of Innovative Optical Health Sciences 2024年第2期17卷 45-58页

作者： Xiaoyan Li Shijie Tu Yile Sun Yubing Han Xiang Hao Cuifang kuang Xu Liu State Key Laboratory of Extreme Photonics and Instrumentation College of Optical Science and Engineering Zhejiang University Hangzhou 310027P.R.China ZJU-Hangzhou Global Scientific and Technological Innovation Center Hangzhou 311200 P.R.China Collaborative Innovation Center of Extreme Optics Shanxi University TaiyuanShanxi 030006P.R.China A dvanced Biomedical Imaging Facility-Wuhan National Laboratory for Optoelectronics Huazhong University of Science and Technology WuhanHubei 430074P.R.China

structured illumination microscopy(SIM)achieves super-resolution(SR)by modulating the high-frequency information of the sample into the passband of the optical system and subsequent image reconstruction.The traditional Wiener-filtering-based reconstruction algorithm operates in the Fourier domain,it requires prior knowledge of the sinusoidal illumination patterns which makes the time-consuming procedure of parameter estimation to raw datasets necessary,besides,the parameter estimation is sensitive to noise or aberration-induced pattern distortion which leads to reconstruction artifacts.Here,we propose a spatial-domain image reconstruction method that does not require parameter estimation but calculates patterns from raw datasets,and a reconstructed image can be obtained just by calculating the spatial covariance of differential calculated patterns and differential filtered datasets(the notch filtering operation is performed to the raw datasets for attenuating and compensating the optical transfer function(OTF)).Experiments on reconstructing raw datasets including nonbiological,biological,and simulated samples demonstrate that our method has SR capability,high reconstruction speed,and high robustness to aberration and noise.

关键词： structured illumination microscopy image reconstruction spatial domain digital micromirror device(DMD)

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

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Advanced Photonics 2023年第5期5卷 96-105页

作者： Yile Sun Hongfei Zhu Lu Yin Hanmeng Wu Mingxuan Cai Weiyun Sun Yueshu Xu Xinxun Yang Jiaxiao Han Wenjie Liu Yubing Han Xiang Hao Renjie Zhou Cuifang Kuang Xu Liu Zhejiang University College of Optical Science and EngineeringState Key Laboratory of Extreme Photonics and InstrumentationHangzhouChina The Chinese University of Hong Kong Department of Biomedical EngineeringHong KongChina China Jiliang University College of Optical and Electronic TechnologyHangzhouChina Zhejiang University of Technology Institute of PharmacologyCollege of Pharmaceutical SciencesHangzhouChina ZJU-Hangzhou Global Scientific and Technological Innovation Center HangzhouChina Huazhong University of Science and Technology Britton Chance Center for Biomedical Photonics-MoE Key Laboratory for Biomedical PhotonicsAdvanced Biomedical Imaging Facility-Wuhan National Laboratory for OptoelectronicsWuhanChina Shanxi University Collaborative Innovation Center of Extreme OpticsTaiyuanChina

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.

关键词： optical imaging super-resolution microscopy fluorescence interference structured illumination microscopy

Improvement in Resolution of Multiphoton Scanning structured illumination microscopy via Harmonics

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Engineering 2022年第9期16卷 65-72页

作者： Lei Wang Xiaomin Zheng Jie Zhou Meiting Wang Jiajie Chen Youjun Zeng Gaixia Xu Ying Wang Haixia Qiu Yonghong Shao Junle Qu Bruce Zhi Gao Ying Gu College of Physics and Optoelectronic Engineering Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong ProvinceShenzhen UniversityShenzhen 518060China Department of Laser Medicine First Medical Center of PLA General HospitalBeijing 100853China Department of Bioengineering and COMSET Clemson UniversityClemsonSC 29634USA

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.

关键词： Super-resolution microscopy structured illumination microscopy Multiphoton-structured illumination microscopy(SIM) SIM Second-harmonic-generation(SHG)-SIM

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

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创新光学健康科学杂志(英文) 2023年第4期16卷 134-149页

作者： Wenshuai Wu Jiajie Chen Meiting Wang Lei Wang Xiaomin Zheng Jia Li Junle Qu Bruce Zhi Gao Yonghong Shao College of Physics and Optoelectronic Engineering Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province Shenzhen University Shenzhen 518060GuangdongP.R.China Department of Bioengineering and COMSET Clemson University ClemsonSC 29634 USA

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.

关键词： Super-resolution structured illumination microscopy second harmonic generation

Nonlinear scanning structured illumination microscopy based on nonsinusoidal modulation

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Journal of Innovative Optical Health Sciences 2021年第5期4卷 25-32页

作者： Meiting Wang Lei Wang Xiaomin Zheng Jie Zhou Jiajie Chen Youjun Zeng Junle Qu Yonghong Shao Bruce Zhi Gao College of Physics and Optoelectronic Engineering Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong ProvinceShenzhen UniversityShenzhen 518060China Department of Bioengineering and COMSET Clemson UniversityClemsonSC 29634USA

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.

关键词： Super-resolution image structured illumination microscopy nonsinusoidal function

structured illumination microscopy imaging of mitochondria in living cells

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Structured illumination microscopy imaging of mitochondria i...

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

第三届中国生物物理学会代谢生物学分会学术研讨会

作者： Xiaoshuai Huang Junchao Fan Liuju Li Shan Tan Liangyi Chen Institute of Molecular Medicine Peking University School of Automation Huazhong University of Science and Technology

1.Introduction The mitochondrion is an essential organelle to produce energy in cells.It is challenging for fluorescence microscopy due to the highly folded inner membrane with topologically complicated structures.structured illumination microscopy(SIM) with our proposed reconstruction method is the most suitable fluorescence microscopy to visualize mitochondrial inner membrane dynamics in living cells owing to a less photodamage and a higher resolution.2.Results The Hessian-SIM can retain the details and suppress the artifacts in the reconstruction.Fig.1 shows the mitochondrial inner membrane in living COS-7 cells labeled with MitoTracker Green FM under wide field(a) and 2 D-SIM(b).It provides a detailed view on the inner membrane architecture in mitochondria.3.Conclusion We proposed a modified parameter estimation method and Hessian-SIM microscopy,which can visualize vivid dynamics of mitochondrial cristae structures during fusion,fission and inter-cristae remodeling in living cells.

关键词： structured illumination microscopy Fast Long-term

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Chinese Chemical Letters 2022年第4期33卷 1865-1869页

作者： Huimin Chen Han Wang Yongchun Wei Maomao Hu Bo Dong Hongbao Fang Qixin Chen Institute of Materia Medica Shandong First Medical University&Shandong Academy of Medical SciencesJi’nan 250062China Department of Cardiology Shandong Provincial Hospital Affiliated to Shandong UniversityJi’nan 250012China State Key Laboratory of Coordination Chemistry Chemistry and Biomedicine Innovation Center(ChemBIC)School of Chemistry and Chemical EngineeringNanjing UniversityNanjing 210023China

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.

关键词： Dextran Subcellular distribution Super-resolution imaging structured illumination microscopy

Application of super-resolution fluorescence microscopy in hematologic malignancies

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Journal of Innovative Optical Health Sciences 2022年第2期15卷 18-31页

作者： Yalan Yu Jianing Yu Zhen-Li Huang Fuling Zhou Department of Hematology Zhongnan Hospital of Wuhan UniversityWuhan 430071China Key Laboratory of Biomedical Engineering of Hainan Province School of Biomedical EngineeringHainan UniversityHaikou 570228China

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.

关键词： Hematologic malignancies super-resolution°uorescence microscopy structured illumination microscopy stimulated emission depletion microscopy single molecule localization microscopy

Super-resolution quantification of nanoscale damage to mitochondria in live cells

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Nano Research 2020年第8期13卷 2149-2155页

作者： Xintian Shao Qixin Chen Lianting Hu Zhiqi Tian Liuyi Liu Fei Liu Fengshan Wang Peixue Ling Zong-Wan Mao Jiajie Diao Shandong Academy of Pharmaceutical Sciences Key Laboratory of BiopharmaceuticalsEngineering Laboratory of Polysaccharide DrugsNational-Local Joint Engineering Laboratory of Polysaccharide DrugsJinan250101China Department of Cancer Biology University of Cincinnati College of MedicineCincinnatiOH45267USA School of Pharmaceutical Sciences Cheeloo College of MedicineShandong UniversityJinan250012China Xintian Shao Qixin Chenand Lianting Hu contributed equally to this work.Shandong First Medical University&Shandong Academy of Medical SciencesJinan250062China Xintian Shao Qixin Chenand Lianting Hu contributed equally to this work.School of Information ManagementWuhan UniversityWuhan430072China MOE Key Laboratory of Bioinorganic and Synthetic Chemistry School of ChemistrySun Yat-Sen UniversityGuangzhou510275China

Mitochondrial damage,characterized by altered morphological distribution and the damage of cristae,is closely associated with mitochondrial disease.However,imaging methods for capturing mitochondrial morphology at the nanoscale level in live samples remain unavailable,which seriously hinders the accurate evaluation and diagnosis of mitochondrial-related diseases.In response,we propose a super-resolution quantification strategy based on structured illumination microscopy(SIM)for the rapid,accurate evaluation of mitochondrial morphology.Using the strategy,we accurately captured the morphological distribution of mitochondria at the nanoscale level in a way generally applicable to checking various cell processes and identifying patients with mitochondrial disease who exhibit the SLC25A46 mutation.We also used algorithm-assisted super-resolution imaging to quantitatively analyze damage to mitochondrial cristae,which supports a novel drug screening strategy—high-resolution drug screening—for investigating drugs’pharmacodynamics on organelles in living cells.In short,our strategy improves the accurate examination of changes in mitochondrial morphology in living cells and indicates new ways in which SIM-imaging can assist in diagnosing mitochondrial disease at the single-cell level.

关键词： mitochondria structured illumination microscopy quantification analyze morphology cristae