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3D-STORM超分辨成像中单分子轴向定位精度优化研究

Optimization of Single Molecules Axial Location Precision in 3D Stochastic Optical Reconstruction Microscopy

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摘要

为定量描述柱透镜参数对三维成像过程中荧光分子轴向定位精度的影响,本文研究了柱透镜参数与点扩散函数椭圆率的相互关系.基于Olympus IX-83倒置荧光显微镜搭建成像系统,利用3个不同焦距柱透镜进行实验,通过柱透镜标定曲线的线性变化范围及该范围内曲线的斜率分析柱透镜参数选择上的优劣,优化并提高轴向定位的精度和深度.选择合适的柱透镜参数可实现标定曲线在焦平面上下1 μm范围线性变化,轴向定位精度为16 nm,并采用优化的标定曲线对肌动蛋白微丝进行三维超分辨成像.

Abstract

The relationship between the ellipticity of point spread function and the focal length of cylindrical lens was investigated. Fluorescent inverted microscopey imaging system is based on Olympus IX-83. With three different focal length,the point spread function with custom built stochastic optical reconstruction microscopy instruments was measured. A method to evaluate the cylindrical lens was developed based on linear region and localization error. The results show that linear region of 1μm and axial localization error of 16 nm can be achieved with correct focal length. As the demonstration,three dimension super-resolution image of Actin filaments was reconstructed.

Newport宣传-MKS新实验室计划
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中图分类号:Q-336

DOI:10.3788/gzxb20154410.1017003

基金项目:国家重大科研仪器研制项目(No.ZDYZ2013-1)、基金委面上项目(No.61475185)和中科院百人计划资助

收稿日期:2015-06-01

修改稿日期:2015-07-27

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作者单位    点击查看

张世超:中国科学院苏州生物医学工程技术研究所 江苏省医用光学重点实验室,江苏 苏州 215163中国科学院长春光学精密机械与物理研究所,长春 130033中国科学院大学,北京 100039
李思黾:中国科学院苏州生物医学工程技术研究所 江苏省医用光学重点实验室,江苏 苏州 215163
杨光:中国科学院苏州生物医学工程技术研究所 江苏省医用光学重点实验室,江苏 苏州 215163
李辉:中国科学院苏州生物医学工程技术研究所 江苏省医用光学重点实验室,江苏 苏州 215163
熊大曦:中国科学院苏州生物医学工程技术研究所 江苏省医用光学重点实验室,江苏 苏州 215163

联系人作者:张世超(1017114279@qq.com)

备注:张世超(1989-),男,硕士研究生,主要研究方向为超分辨率显微成像技术.

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引用该论文

ZHANG Shi-chao,LI Si-min,YANG Guang,LI Hui,XIONG Da-xi. Optimization of Single Molecules Axial Location Precision in 3D Stochastic Optical Reconstruction Microscopy[J]. ACTA PHOTONICA SINICA, 2015, 44(10): 1017003

张世超,李思黾,杨光,李辉,熊大曦. 3D-STORM超分辨成像中单分子轴向定位精度优化研究[J]. 光子学报, 2015, 44(10): 1017003

被引情况

【1】吴瑞祥,陈瑞云,乔志星,高岩,张国峰,秦成兵,肖连团,贾锁堂. 氧气诱导单分子电子转移的动力学特性. 激光与光电子学进展, 2017, 54(5): 53004--1

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