Periodic acceptor excitation spectroscopy of single molecules

被引：15
作者：
Doose, Soren
Heilemann, Mike
Michalet, Xavier
Weiss, Shimon
Kapanidis, Achillefs N. [1 ]
机构：
[1] Univ Bielefeld, Appl Laser Phys & Laser Spectroscopy, D-4800 Bielefeld, Germany
[2] Univ Oxford, IRC Bionanotechnol, Dept Phys, Clarendon Lab, Oxford, England
[3] Univ Calif Los Angeles, Dept Biochem & Chem, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, Dept Physiol, Los Angeles, CA 90095 USA
来源：
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 2007年 / 36卷 / 06期
基金：
英国工程与自然科学研究理事会;
关键词：
single-molecule fluorescence spectroscopy; alternating-laser excitation (ALEX); Forster resonance energy transfer (FRET); biomolecular interactions;
D O I：
10.1007/s00249-007-0133-7
中图分类号：
Q6 [生物物理学];
学科分类号：
071011 ;
摘要：
Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Forster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.
引用
收藏
页码：669 / 674
页数：6
相关论文
共 18 条
[1]   Separating structural heterogeneities from stochastic variations in fluorescence resonance energy transfer distributions via photon distribution analysis [J].
Antonik, M ;
Felekyan, S ;
Gaiduk, A ;
Seidel, CAM .
JOURNAL OF PHYSICAL CHEMISTRY B, 2006, 110 (13) :6970-6978
[2]   Short-range spectroscopic ruler based on a single-molecule optical switch [J].
Bates, M ;
Blosser, TR ;
Zhuang, XW .
PHYSICAL REVIEW LETTERS, 2005, 94 (10)
[3]   Single-pair fluorescence resonance energy transfer on freely diffusing molecules: Observation of Forster distance dependence and subpopulations [J].
Deniz, AA ;
Dahan, M ;
Grunwell, JR ;
Ha, TJ ;
Faulhaber, AE ;
Chemla, DS ;
Weiss, S ;
Schultz, PG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (07) :3670-3675
[4]   Comparison of photophysical and colloidal properties of biocompatible semiconductor nanocrystals using fluorescence correlation spectroscopy [J].
Doose, S ;
Tsay, JM ;
Pinaud, F ;
Weiss, S .
ANALYTICAL CHEMISTRY, 2005, 77 (07) :2235-2242
[5]   Single-molecule fluorescence resonance energy transfer [J].
Ha, T .
METHODS, 2001, 25 (01) :78-86
[6]   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
[7]   Retention of transcription initiation factor σ70 in transcription elongation:: Single-molecule analysis [J].
Kapanidis, AN ;
Margeat, E ;
Laurence, TA ;
Doose, S ;
Ho, SO ;
Mukhopadhyay, J ;
Kortkhonjia, E ;
Mekler, V ;
Ebright, RH ;
Weiss, S .
MOLECULAR CELL, 2005, 20 (03) :347-356
[8]   Fluorescence-aided molecule sorting: Analysis of structure and interactions by alternating-laser excitation of single molecules [J].
Kapanidis, AN ;
Lee, NK ;
Laurence, TA ;
Doose, S ;
Margeat, E ;
Weiss, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (24) :8936-8941
[9]   Alternating-laser excitation of single molecules [J].
Kapanidis, AN ;
Laurence, TA ;
Lee, NK ;
Margeat, E ;
Kong, XX ;
Weiss, S .
ACCOUNTS OF CHEMICAL RESEARCH, 2005, 38 (07) :523-533
[10]   A single-molecule Forster resonance energy transfer analysis of fluorescent DNA-protein conjugates for nanobiotechnology [J].
Kukolka, Florian ;
Mueller, Barbara K. ;
Paternoster, Stefan ;
Arndt, Andreas ;
Niemeyer, Christof M. ;
Braeuchle, Christoph ;
Lamb, Don C. .
SMALL, 2006, 2 (8-9) :1083-1089
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