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Super-resolution microscopy of live cells using single molecule localization

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Chinese Science Bulletin 2013年第36期58卷 4519-4527页

作者： ZHANG YongDeng CHANG Hao gu lusheng ZHAO YanHua XU Tao JI Wei School of Life Science and Technology Huazhong University of Science and Technology School of Life Science University of Science and Technology of China Institute of Biophysics Chinese Academy of Sciences

The resolution of conventional light microscopy is insufficient for subcelluar *** invention of various super-resolution imaging techniques breaks the diffraction barrier and pushes the resolution limit towards the nanometer ***,we focus on a category of super-resolution microscopy that relies on the stochastic activation and precise localization of single molecules.A diversity of fluorescent probes with different characteristics has been developed to achieve super-resolution *** addition,with the implementation of robust localization algorithms,this family of approaches has been expanded to multi-color,three-dimensional and live cell imaging,which provides a promising prospect in biological research.

关键词：超高分辨率光学显微镜活细胞成像分子定位成像技术超分辨率分辨率极限纳米尺度

Targeted disruption of RablO causes early embryonic lethality

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Protein & Cell 2015年第6期6卷 463-467页

作者： Pingping Lv Yi Sheng Zhenao Zhao Wei Zhao lusheng gu Tao Xu Eli Song National Laboratory of Biomacromolecules institute of Biophysics Chinese Academy of Sciences Beiiin~l 100101 China University of Chinese Academy of Sciences Beijing 100049 China College of Life Science and Technology Huazhong University of Science and Technology Wuhan 430074 China State Key Laboratory of Reproductive Biology Institute of Zoology Chinese Academy of Sciences Beijing 100101 China

Dear Editor, Intracellular trafficking is a basis of cellular activities, including cell migration, immune response, and development （Lebreton et al., 2003; Lu（;in et al., 2014; Ulrich and Heisenberg, 2009）. In healthy tissue, intracellular trafficking is highly regulated, con- trolling the form and function of cells （Furthauer and Gonzalez- Gaitan, 2009）.

关键词： Early Endosome Mutant Embryo Recycling Endosome Massive Cell Death Endosomal Trafficking

Ultra-stable super-resolution fluorescence cryo-microscopy for correlative light and electron cryo-microscopy

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Science China(Life Sciences) 2018年第11期61卷 1312-1319页

作者： Xiaojun Xu Yanhong Xue Buyun Tian Fengping Feng lusheng gu Weixing Li Wei Ji Tao Xu College of Life Science and Technology Huazhong University of Science and TechnologyWuhan 430074China National Laboratoty of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China Center for Biological Instrument Development Core Facility for Protein ResearchInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China College of Life Science University of Chinese Academy of SciencesBeijing 100049 China

Remarkable progress in correlative light and electron cryo-microscopy(cryo-CLEM) has been made in the past decade. A crucial component for cryo-CLEM is a dedicated cryo-fluorescence microscope(cryo-FM). Here, we describe an ultra-stable superresolution cryo-FM that exhibits excellent thermal and mechanical stability. The temperature fluctuations in 10 h are less than0.06 K, and the mechanical drift over 5 h is less than 200 nm in three dimensions. We have demonstrated the super-resolution imaging capability of this system(average single molecule localization accuracy of ～13.0 nm). The results suggest that our system is particularly suitable for long-term observations, such as single molecule localization microscopy(SMLM) and cryogenic super-resolution correlative light and electron microscopy(csCLEM).

关键词： cryogenic cryostat super-resolution Cryo-FM Cryo-PALM csCLEM

CAMSAP2 and CAMSAP3 localize at microtubule intersections to regulate the spatial distribution of microtubules

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Journal of Molecular Cell Biology 2023年第8期15卷 28-39页

作者： Rui Zhang lusheng gu Wei Chen Nobutoshi Tanaka Zhengrong Zhou Honglin Xu Tao Xu Wei Ji Xin Liang Wenxiang Meng State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeĳing 100101China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeĳing 100049China National Laboratory of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeĳing 100101China Innovation Center of Optical Imaging and Detection Technology R&D Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory)Guangzhou 510320China IDG/McGovern Institute for Brain Research Tsinghua–Peking Joint Center for Life ScienceSchool of Life SciencesTsinghua UniversityBeĳing 100084China Laboratory for Cell Adhesion and Tissue Patterning RIKEN Center for Developmental Biology/RIKEN Center for Biosystems Dynamics Research650-0047 KobeJapan Innovation Academy for Seed Design Chinese Academy of SciencesBeĳing 100101China

Microtubule networks support many cellular processes and exhibit a highly ordered ***,due to the limited axial resolution of conventional light microscopy,the structural features of these networks cannot be resolved in three-dimensional(3D)***,we used customized ultra-high-resolution interferometric single-molecule localization microscopy to characterize the microtubule networks in Caco2 *** found that the calmodulin-regulated spectrin-associated proteins(CAMSAPs)localize at a portion of microtubule *** investigation showed that depletion of CAMSAP2 and CAMSAP3 leads to the narrowing of the inter-microtubule ***,CAMSAPs recognize microtubule defects,which often occur near microtubule intersections,and then recruit katanin to remove the damaged ***,the CAMSAP–katanin complex is a regulatory module for the distance between *** together,our results characterize the architecture of cellular microtubule networks in high resolution and provide molecular insights into how the 3D structure of microtubule networks is controlled.

关键词： CAMSAPs microtubule intersections microtubule networks

Ultrafast,accurate,and robust localization of anisotropic dipoles

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Protein & Cell 2013年第8期4卷 598-606页

作者： Yongdeng Zhang lusheng gu Hao Chang Wei Ji Yan Chen Mingshu Zhang Lu Yang Bei Liu Liangyi Chen Tao Xu College of Life Science and Technology Huazhong University of Science and TechnologyWuhan 430074China National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China School of Life Sciences University of Science and Technology of ChinaHefei 230026China University of Chinese Academy of Sciences Beijing 100049China Laboratory of Cell Secretion and Metabolism Institute of Molecular MedicinePeking UniversityBeijing 100871China

The resolution of single molecule localization imaging techniques largely depends on the precision of localization ***,the commonly used Gaussian function is not appropriate for anisotropic dipoles because it is not the true point spread *** derived the theoretical point spread function of tilted dipoles with restricted mobility and developed an algorithm based on an artifi cial neural network for estimating the localization,orientation and mobility of individual *** with fi tting-based methods,our algorithm demonstrated ultrafast speed and higher accuracy,reduced sensitivity to defocusing,strong robustness and adaptability,making it an optimal choice for both two-dimensional and threedimensional super-resolution imaging analysis.

关键词： point spread function restricted mobility artifi cial neural network super-resolution imaging

Spatial distribution of IL4 controls iNKT cell-DC crosstalk in tumors

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Cellular & Molecular Immunology 2020年第5期17卷 496-506页

作者： Lu Wang Zhilan Liu Lili Wang Qielan Wu Xiang Li Di Xie Huimin Zhang Yongdeng Zhang lusheng gu Yanhong Xue Ting Yue Gang Liu Wei Ji Haiming Wei Tao Xu Li Bai Division of Life Sciences and Medicine Department of Oncology of The First Affiliated HospitalThe CAS Key Laboratory of Innate Immunity and Chronic DiseaseUniversity of Science and Technology of China230027 HefeiChina National Key Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of Sciences100101 BeijingChina School of Life Sciences University of Science and Technology of China230027 HefeiChina National Synchrotron Radiation Laboratory University of Science and Technology of China230027 HefeiChina

The spatiotemporal distribution of cytokines orchestrates immune responses in vivo,yet the underlying mechanisms remain to be *** showed here that the spatial distribution of interleukin-4(IL4)in invariant natural killer T(iNKT)cells regulated crosstalk between iNKT cells and dendritic cells(DCs)and controlled iNKT cell-mediated T-helper type 1(Th1)*** persistent polarization of IL4 induced by strong lipid antigens,that is,α-galactosylceramide(αGC),caused IL4 accumulation at the immunological synapse(IS),which promoted the activation of the IL4R-STAT6(signal transducer and activator of transcription 6)pathway and production of IL12 in DCs,which enhanced interferon-γ(IFNγ)production in iNKT ***,the nonpolarized secretion of IL4 induced by Th2 lipid antigens with a short or unsaturated chain was incapable of enhancing this iNKT cell-DC crosstalk and thus shifted the immune response to a Th2-type *** nonpolarized secretion of IL4 in response to Th2 lipid antigens was caused by the degradation of Cdc42 in iNKT ***,reduced Cdc42 expression was observed in tumorinfiltrating iNKT cells,which impaired IL4 polarization and disturbed iNKT cell-DC crosstalk in tumors.

关键词： Spatial distribution Polarization IL4 iNKT DC Crosstalk Tumor

Tetra-color superresolution microscopy based on excitation spectral demixing

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Light(Science & Applications) 2023年第1期12卷 99-108页

作者： Wanyan Wu Shihang Luo Chunyan Fan Tianjie Yang Shuwen Zhang Wenxiang Meng Tao Xu Wei Ji lusheng gu Institute of Biophysics Chinese Academy of SciencesBeijing 100101China College of Life Science University of Chinese Academy of SciencesBeijing 100049China State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China School of Future Technology University of Chinese Academy of SciencesBeijing 100049China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing 100049China Bioland Laboratory Guangzhou 510005China guangzhou Laboratory Guangzhou 510030China

Multicolor imaging allows protein colocalizations and organelle interactions to be studied in biological research,which is especially important for single-molecule localization microscopy(SMLM).Here,we propose a multicolor method called excitation-resolved stochastic optical reconstruction microscopy(ExR-STORM).The method,which is based on the excitation spectrum of fluorescent dyes,successfully separated four spectrally very close far-red organic fluorophores utilizing three excitation lasers with cross-talk of less than 3%.Dyes that are only 5 nm apart in the emission spectrum were resolved,resulting in negligible chromatic *** method was extended to three-dimensional(3D)imaging by combining the astigmatic method,providing a powerful tool for resolving 3D morphologies at the nanoscale.

关键词： excitation spectral mixing

The Inherent Law of the Unpredictability of Financial Asset Price Fluctuations: Multistability and Chaos

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Journal of Systems Science & Complexity 2024年第2期37卷 776-804页

作者： gu Enguo Huanghe University of Science and Technology Zhengzhou 450006China

This paper aims at understanding the price dynamics generated by the interaction of traders relying on heterogeneous expectations in an asset pricing *** the present work the authors analyze a financial market populated by five types of boundedly rational speculators-two types of fundamentalists,two types of chartists and trend followers which submit buying/selling orders according to different trading *** authors formulate a stock market model represented as a 2 dimensional piecewise linear discontinuous *** proposed contribution to the existing financial literature is two ***,the authors perform study of the model involving a 2 dimensional piecewise linear discontinuous map through a combination of qualitative and quantitative *** authors focus on the existence conditions of chaos and the multi-stability regions in parameter *** border collision bifurcation curves and basins of multi-attractors are also *** authors find that chaos or quasi-period exists only in the case of fixed point being a saddle(regular or flip)and that the coexistence of multiple attractors may exist when the fixed point is an attractor,but it is common for spiral and flip fixed points.

关键词： BCB curve chaos financial market heterogeneous agents multiple attractors piecewise linear discontinuous map

Enhanced remediation of chlorpyrifos-contaminated soil by immobilized strain Bacillus H27

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Journal of Environmental Sciences 2024年第10期144卷 172-184页

作者： Changrui Liu Shengfang Wen Shuhan Li Yu Tian Lanjun Wang lusheng Zhu Jun Wang Young Mo Kim Jinhua Wang National Engineering Research Center for Efficient Utilization of Soil and Fertilizer Resources Key Laboratory of Agricultural Environment in Universities of ShandongCollege of Resources and EnvironmentShandong Agricultural UniversityTaian 271018China Department of Civil and Environmental Engineering Hanyang UniversitySeongdong-guSeoul 04763Korea

Chlorpyrifos is a pesticide widely used in agricultural production with a relatively long residual half-life in *** the problem of residual chlorpyrifos is of universal *** this study,rice hull biochar was used as an immobilized carrier to prepare the immobilized strain H27 for the remediation of chlorpyrifos-contamination *** microorganisms after remediation were investigated by ecotoxicological *** immobilized strain H27 had the highest removal rate of chlorpyrifos when 10%bacterial solution was added to the liquid medium containing 0.075-0.109 mm diameter biochar cultured for 22 *** study on the removal of chlorpyrifos by immobilized strain H27 showed that the initial concentration of chlorpyrifos in solution was 25mg/L,and the removal rate reached 97.4%after 7 days of *** the soil,the removal rate of the immobilized bacteria group increased throughout the experiment,which was significantly higher than that of the free bacteria and biochar treatment *** Biolog-ECO test,T-RFLP and RT-RCR were used to study the effects of the soil microbial community and nitrogen cycling functional genes during chlorpyrifos *** was found that ICP group had the highest diversity index among the four treatment *** microflora of segment containing 114 bp was the dominant bacterial community,and the dominant microflora of the immobilized bacteria group was more evenly *** influence of each treatment group on ammonia-oxidizing bacteria(AOB)was greater than on ammonia-oxidizing archaea(AOA).This study offers a sound scientific basis for the practical application of immobilized bacteria to reduce residual soil pesticides.

关键词： Chlorpyrifos Immobilized microorganisms Soil remediation Soil microorganism