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利用衍射光学元件调制轴对称偏振光提高荧光显微镜纵向分辨率

Improve Axial Resolution of Fluorescent Microscopy Using Cylindrical Vector Beams Modulated by Diffractive Optical Elements

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摘要

利用荧光蛋白激活状态的可逆特性,提出了一种提高荧光显微镜纵向分辨率的方法。针对不同数值孔径的显微系统,通过合理选择带涡旋相位的轴对称偏振光作为激活光和退活光,并设计衍射光学元件对其进行调制,从而实现百纳米级的纵向选择激活。而且,通过调节激活光和退活光的光强比率R,可以控制荧光蛋白最大激活概率满足单分子荧光显微技术的需求,同时进一步降低纵向选择激活层半峰全宽(FWHM)至25.7 nm,并且约90%的激活荧光蛋白位于30 nm厚的激活层中(R=500)。

Abstract

A novel method to improve the axial resolution of fluorescent microscopy using photo-switching characteristics of fluorescent protein is proposed. For microscopy with different numerical apertures, a 100-nm axially-selective-activation is achieved by using cylindrical vector beams with vortex phase as activation and deactivation beams which are modulated by designed diffractive optical elements. Furthermore, adjusting the power ratio R of the activation beam and deactivation beam will make the maximum activation probability of fluorescent protein suitable for single-molecule microscopy. The numerical calculation based on Cy5 demonstrates that along the z axis the activation probability has a single peak with full-width at half-maximum (FWHM) of only 25.7 nm and about 90% of activated molecules are located in a 30-nm-thick layer around zero point (for R=500).

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中图分类号:O436.1

DOI:10.3788/cjl201340.0410001

所属栏目:微纳光学

责任编辑:李文喆  信息反馈

收稿日期:2012-12-10

修改稿日期:2012-12-18

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作者单位    点击查看

曹国威:中国电子科技集团第三十八研究所光电集成研究中心, 安徽 合肥 230088
李永平:中国科学技术大学物理学院, 安徽 合肥 230026
毕国强:中国科学技术大学生命科学学院, 安徽 合肥 230026

联系人作者:曹国威(cgw2003@mail.ustc.edu.cn)

备注:曹国威(1987—),男,博士,工程师,主要从事衍射光学元件方面的研究。

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引用该论文

Cao Guowei,Li Yongping,Bi Guoqiang. Improve Axial Resolution of Fluorescent Microscopy Using Cylindrical Vector Beams Modulated by Diffractive Optical Elements[J]. Chinese Journal of Lasers, 2013, 40(4): 0410001

曹国威,李永平,毕国强. 利用衍射光学元件调制轴对称偏振光提高荧光显微镜纵向分辨率[J]. 中国激光, 2013, 40(4): 0410001

被引情况

【1】冀胜哲,朱林伟,孙美玉,王丽丽,李志刚. 紧聚焦条件下的轴向双焦点波带片. 光学学报, 2015, 35(3): 305003--1

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