A 3D Aligning Method for Stimulated Emission Depletion Microscopy Using Fluorescence Lifetime Distribution

被引：10

作者：

Wang, Yifan ^{[1
]}

Kuang, Cuifang ^{[1
]}

Li, Shuai ^{[1
]}

Hao, Xiang ^{[1
]}

Xu, Yingke ^{[2
]}

Liu, Xu ^{[1
]}

机构：

[1] Zhejiang Univ, State Key Lab Modern Opt Instrumentat, Dept Opt Engn, Hangzhou 310027, Peoples R China

[2] Zhejiang Univ, Dept Biomed Engn, Minist Educ, Key Lab Biomed Engn, Hangzhou 310027, Peoples R China

来源：

MICROSCOPY RESEARCH AND TECHNIQUE | 2014年 / 77卷 / 11期

基金：

中国国家自然科学基金;

关键词：

aligning method; STED; fluorescence microscopy; LIMIT;

D O I：

10.1002/jemt.22420

中图分类号：

R602 [外科病理学、解剖学]; R32 [人体形态学];

学科分类号：

100101 ;

摘要：

Optimal resolution by stimulated emission depletion (STED) microscopy requires precise alignment of the donut-shaped depletion focus to the excitation focus. In this article, we demonstrate that fluorescence lifetime distribution can be implemented to align the STED system. Different from the traditional aligning methods in which a scattering imaging module is often equipped, the lifetime-based method is free from probable mismatches between the scattering mode and the fluorescent mode, drift errors caused by separate imaging and complex fitting methods. Based on this method, a spatial resolution of 38 nm by time-gated detection has been achieved. Microsc. Res. Tech. 77:935-940, 2014. (c) 2014 Wiley Periodicals, Inc.

引用

页码：935 / 940

页数：6

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