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Single-cell manipulation by two-dimensional micropatterning
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作者 Xuehe Ma Haimei Zhang +7 位作者 Shiyu Deng Qiushuo Sun Qingsong Hu Yuhang Pan Fen Hu imshik lee Fulin Xing Leiting Pan 《Journal of Innovative Optical Health Sciences》 SCIE EI CSCD 2024年第1期45-59,共15页
Cells are highly sensitive to their geometrical and mechanical microenvironment that directly regulate cell shape,cytoskeleton and organelle,as well as the nucleus morphology and genetic expression.The emerging two-di... Cells are highly sensitive to their geometrical and mechanical microenvironment that directly regulate cell shape,cytoskeleton and organelle,as well as the nucleus morphology and genetic expression.The emerging two-dimensional micropatterning techniques offer powerful tools to construct controllable and well-organized microenvironment for single-cell level investigations with qualitative analysis,cellular standardization,and in vivo environment mimicking.Here,we provide an overview of the basic principle and characteristics of the two most widely-used micropatterning techniques,including photolithographic micropatterning and soft lithography micropatterning.Moreover,we summarize the application of micropatterning technique in controlling cytoskeleton,cell migration,nucleus and gene expression,as well as intercellular communication. 展开更多
关键词 Two-dimensional micropatterning CYTOSKELETON cell migration extracellular matrix intercellular communication gene expression
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Regulation of actin cytoskeleton via photolithographic micropatterning 被引量:2
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作者 Fulin Xing Haimei Zhang +7 位作者 Mengyu Li Hao Dong Xuehe Ma Shiyu Deng Fen Hu imshik lee Leiting Pan Jingjun Xu 《Journal of Innovative Optical Health Sciences》 SCIE EI CAS CSCD 2023年第2期50-57,共8页
Actin cytoskeleton plays crucial roles in various cellular functions.Extracellular matrix(ECM)can modulate cell morphology by remodeling the internal cytoskeleton.To define how geometry of ECM regulates the organizati... Actin cytoskeleton plays crucial roles in various cellular functions.Extracellular matrix(ECM)can modulate cell morphology by remodeling the internal cytoskeleton.To define how geometry of ECM regulates the organization of actin cytoskeleton,we plated individual NIH 3T3 cells on micropatterned substrates with distinct shapes and sizes.It was found that the stress fibers could form along the nonadhesive edges of T-shaped pattern,but were absent from the opening edge of V-shaped pattern,indicating that the organization of actin cytoskeleton was dependent on the mechanical environment.Furthermore,a secondary actin ring was observed on 50μm circular pattern while did not appear on 30μm and 40μm pattern,showing a size-dependent organization of actin cytoskeleton.Finally,osteoblasts,MDCK and A549 cells exhibited distinct organization of actin cytoskeleton on T-shaped pattern,suggesting a cell-type specificity in arrangement of actin cytoskeleton.Together,our findings brought novel insight into the organization of actin cytoskeleton on micropatterned environments. 展开更多
关键词 Actin cytoskeleton PHOTOLITHOGRAPHY MICROPATTERNING extracellular matrix
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Structure Studies on Host-vip Recognition Sensory Systems
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作者 Lin Jing YANG Xi Zeng FENG +1 位作者 imshik lee Chun Li BAI(Institute of Chemistry, the Chinese Academy of Science,Beijing, 100080) 《Chinese Chemical Letters》 SCIE CAS CSCD 1997年第8期707-710,共4页
Rhodamine B-ethylenediamine-beta-cyclodextrins (RhB-beta-CDen) and rhodamine B-beta-cyclodextrins (RhB-beta-CD) form inclusion complexes with many vip molecules, which can be used as nucleic acid probe. In this pape... Rhodamine B-ethylenediamine-beta-cyclodextrins (RhB-beta-CDen) and rhodamine B-beta-cyclodextrins (RhB-beta-CD) form inclusion complexes with many vip molecules, which can be used as nucleic acid probe. In this paper we determined the most stable conformations of RhB-beta-CDen and RhB-beta-CD by molecular mechanics and dynamics simulation. The interaction between RhB-beta-CDen and two vip molecules, 1-borneol and cyclohexanol, have been investigated both theoretically and experimentally. The results show that the interaction between borneol and RhB-beta-CDen is stronger than that between cyclohexanol and RhB-beta-CDen. 展开更多
关键词 RHB Structure Studies on Host-vip Recognition Sensory Systems
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二抗物种特异性对双色STORM成像的影响(特邀) 被引量:1
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作者 邓师禹 胡芬 +3 位作者 侯梦迪 杨建宇 李任植 潘雷霆 《激光与光电子学进展》 CSCD 北大核心 2024年第6期145-152,共8页
随机光学重建显微术(STORM)基于免疫荧光标记技术,具有原理易懂、光路简单、分辨率极高等特点,一直受到科研工作者的青睐,但分辨率的提升对抗体的特异性提出了更高的要求。相较一抗直接标记,“一抗+二抗”的间接标记法在实际应用中普适... 随机光学重建显微术(STORM)基于免疫荧光标记技术,具有原理易懂、光路简单、分辨率极高等特点,一直受到科研工作者的青睐,但分辨率的提升对抗体的特异性提出了更高的要求。相较一抗直接标记,“一抗+二抗”的间接标记法在实际应用中普适性更强。二抗相对一抗存在物种特异性的问题,生产时需要对其进行预吸附来提升物种特异性。为了探究二抗物种特异性对双色STORM成像的影响,基于经典的红细胞骨架模型中血影蛋白N端和C端的互斥位置关系,对二者使用高、低吸附二抗标记后分别进行双色STORM成像,对照模拟中有无信号串扰条件下的互相关分析结果,结果表明低吸附二抗会造成二者共定位的假象。进一步,分别通过高、低吸附二抗对MDA-MB-231乳腺癌细胞CD47和PD-L1两种膜蛋白进行双色STORM成像,结果揭示两种蛋白无共定位关系。本研究为二抗物种特异性的评估提供了一种基于红细胞骨架结构模型的超分辨成像新策略,助力双色STORM成像精准阐明蛋白分子互作关系。 展开更多
关键词 超分辨成像 随机光学重建显微术 免疫荧光 红细胞膜骨架 互相关分析
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结合多次DBSCAN和层次聚类算法的膜蛋白单分子定位超分辨图像分割 被引量:8
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作者 杨建宇 胡芬 +5 位作者 邢福临 董浩 侯梦迪 李任植 潘雷霆 许京军 《中国激光》 EI CAS CSCD 北大核心 2023年第3期78-85,共8页
膜蛋白在细胞膜上的时空分布形式决定了其活性状态及功能,在调控细胞生命活动过程中起着重要作用。单分子定位超分辨成像(SMLM)技术为在纳米尺度解析膜蛋白的空间分布提供了可能,但分辨率的极大提升对图像准确聚类分割提出了更高要求。... 膜蛋白在细胞膜上的时空分布形式决定了其活性状态及功能,在调控细胞生命活动过程中起着重要作用。单分子定位超分辨成像(SMLM)技术为在纳米尺度解析膜蛋白的空间分布提供了可能,但分辨率的极大提升对图像准确聚类分割提出了更高要求。基于密度的空间聚类算法(DBSCAN)是常用的聚类方法之一,但其对于膜蛋白分布不均匀的SMLM超分辨图像的分割效果往往不太理想。本文提出了一种结合多次DBSCAN和层次聚类的混合聚类算法,该算法以DBSCAN方法为分割基础,通过进一步的面积阈值分析和层次聚类,在保持超分辨点簇图像精确聚类识别的前提下,仍能保留每个点簇内的多次定位信号。将该算法应用于模拟数据集和实验数据分割得到的轮廓系数等性能普遍优于传统DBSCAN算法。这种混合聚类方法为膜蛋白SMLM超分辨图像的聚类分割提供了新思路和新方法,有助于更精准地分析膜蛋白在纳米尺度上的空间分布信息。 展开更多
关键词 生物光学 单分子定位超分辨成像 超分辨图像分割 膜蛋白 基于密度的空间聚类算法 层次聚类算法
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