Nonblinking and longlasting single-molecule fluorescence imaging

被引：508

作者：

Rasnik, Ivan ^{[1
]}

McKinney, Sean A.

Ha, Taekjip

机构：

[1] Emory Univ, Dept Phys, Atlanta, GA 30322 USA

[2] Univ Illinois, Dept Phys, Urbana, IL 61801 USA

[3] Howard Hughes Med Inst, Urbana, IL 61801 USA

来源：

NATURE METHODS | 2006年 / 3卷 / 11期

关键词：

D O I：

10.1038/NMETH934

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Photobleaching and blinking of fluorophores pose fundamental limitations on the information content of single-molecule fluorescence measurements. Photoinduced blinking of Cy5 has hampered many previous investigations using this popular fluorophore. Here we show that Trolox in combination with the enzymatic oxygen-scavenging system eliminates Cy5 blinking, dramatically reduces photobleaching and improves the signal linearity at high excitation rates, significantly extending the applicability of single-molecule fluorescence techniques.

引用

收藏

页码：891 / 893

页数：3

相关论文

共 15 条

[1] Short-range spectroscopic ruler based on a single-molecule optical switch [J].

Bates, M ;

Blosser, TR ;

Zhuang, XW .

PHYSICAL REVIEW LETTERS, 2005, 94 (10)

[2] ENZYMATIC REMOVAL OF OXYGEN FOR POLAROGRAPHY AND RELATED METHODS [J].

BENESCH, RE ;

BENESCH, R .

SCIENCE, 1953, 118 (3068) :447-448

[3] tRNA dynamics on the ribosome during translation [J].

Blanchard, SC ;

Kim, HD ;

Gonzalez, RL ;

Puglisi, JD ;

Chu, S .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (35) :12893-12898

[4] Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time [J].

Churchman, LS ;

Ökten, Z ;

Rock, RS ;

Dawson, JF ;

Spudich, JA .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (05) :1419-1423

[5] Non-exponential bleaching of single bioconjugated Cy5 molecules [J].

Füreder-Kitzmüller, E ;

Hesse, J ;

Ebner, A ;

Gruber, HJ ;

Schütz, GJ .

CHEMICAL PHYSICS LETTERS, 2005, 404 (1-3) :13-18

[6] Monitoring the conformational fluctuations of DNA hairpins using single-pair fluorescence resonance energy transfer [J].

Grunwell, JR ;

Glass, JL ;

Lacoste, TD ;

Deniz, AA ;

Chemla, DS ;

Schultz, PG .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2001, 123 (18) :4295-4303

[7] Single-molecule fluorescence resonance energy transfer [J].

Ha, T .

METHODS, 2001, 25 (01) :78-86

[8] Temporal fluctuations of fluorescence resonance energy transfer between two dyes conjugated to a single protein [J].

Ha, TJ ;

Ting, AY ;

Liang, J ;

Deniz, AA ;

Chemla, DS ;

Schultz, PG ;

Weiss, S .

CHEMICAL PHYSICS, 1999, 247 (01) :107-118

[9] Carbocyanine dyes as efficient reversible single-molecule optical switch [J].

Heilemann, M ;

Margeat, E ;

Kasper, R ;

Sauer, M ;

Tinnefeld, P .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (11) :3801-3806

[10] FORCE MEASUREMENTS BY MICROMANIPULATION OF A SINGLE ACTIN FILAMENT BY GLASS NEEDLES [J].

KISHINO, A ;

YANAGIDA, T .

NATURE, 1988, 334 (6177) :74-76