Autofocusing system based on optical astigmatism analysis of single-molecule images

被引：11

作者：

Hwang, Wonseok ^{[1
,3
]}

Bae, Sangsu ^{[1
,3
]}

Hohng, Sungchul ^{[1
,2
,3
]}

机构：

[1] Seoul Natl Univ, Dept Phys & Astron, Seoul 151747, South Korea

[2] Seoul Natl Univ, Dept Biophys & Chem Biol, Seoul 151747, South Korea

[3] Seoul Natl Univ, Natl Ctr Creat Res Initiat, Seoul 151747, South Korea

来源：

OPTICS EXPRESS | 2012年 / 20卷 / 28期

基金：

新加坡国家研究基金会;

关键词：

CONFORMATIONAL-CHANGES; DRIFT CORRECTION; FRET; MICROSCOPY; FLUORESCENCE; DNA; LOCALIZATION; REGISTRATION;

D O I：

10.1364/OE.20.029353

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

Single-molecule fluorescence imaging has greatly contributed to our understanding of many bio-molecular systems. While reactions occurring in the range of several minutes can be readily studied using conventional single-molecule fluorescence microscopes, data acquisition for longer time scales is hindered by the focal drift of high numerical aperture objectives, which should be corrected in real time. Here, we developed a robust autofocusing system based on optical astigmatism analysis of single-molecule images. Compared to the previously developed methods, our approach has a merit of simplicity in that neither fiducial makers nor an additional laser-detector system is required. As a demonstration, we observed B-Z transition dynamics occurring for several hours. (C) 2012 Optical Society of America

引用

页码：29353 / 29360

页数：8

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