Nanometer-scale mapping and single-molecule detection with color-coded nanoparticle probes

被引:78
作者
Agrawal, Amit [4 ,5 ,6 ]
Deo, Rajesh [3 ]
Wang, Geoffrey D. [4 ,5 ,6 ]
Wang, May D. [1 ,2 ]
Nie, Shuming [4 ,5 ,6 ]
机构
[1] Georgia Inst Technol, Dept Biomed Engn, Atlanta, GA 30332 USA
[2] Emory Univ, Atlanta, GA 30332 USA
[3] Georgia State Univ, Dept Phys & Astron, Atlanta, GA 30303 USA
[4] Emory Univ, Dept Biomed Engn, Atlanta, GA 30322 USA
[5] Emory Univ, Dept Chem, Atlanta, GA 30322 USA
[6] Georgia Inst Technol, Atlanta, GA 30322 USA
关键词
colocalization; multicolor; superresolution;
D O I
10.1073/pnas.0712351105
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We report a method for single-molecule detection and biomolecular structural mapping based on dual-color imaging and automated colocalization of bioconjugated nanoparticle probes at nanometer precision. In comparison with organic dyes and fluorescent proteins, nanoparticle probes such as fluorescence energy-transfer nanobeads and quantum dots provide significant advantages in signal brightness, photostability, and multicolor-light emission. As a result, we have achieved routine two-color super-resolution imaging and single-molecule detection with standard fluorescence microscopes and inexpensive digital color cameras. By using green and red nanoparticles to simultaneously recognize two binding sites on a single target, individual biomolecules such as nucleic acids are detected and identified without target amplification or probe/target separation. We also demonstrate that a powerful astrophysical method (originally developed to analyze crowded stellar fields) can be used for automated and rapid statistical analysis of nanoparticle colocalization signals. The ability to rapidly localize bright nanoparticle probes at nanometer precision has implications not only for ultrasensitive medical detection but also for structural mapping of molecular complexes in which individual components are tagged with color-coded nanoparticles.
引用
收藏
页码:3298 / 3303
页数:6
相关论文
共 46 条
[21]   Ultrahigh-resolution multicolor colocalization of single fluorescent probes [J].
Lacoste, TD ;
Michalet, X ;
Pinaud, F ;
Chemla, DS ;
Alivisatos, AP ;
Weiss, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (17) :9461-9466
[22]   Molecule by molecule direct and quantitative counting of antibody-protein complexes in solution [J].
Li, HT ;
Zhou, DJ ;
Browne, H ;
Balasubramanian, S ;
Klenerman, D .
ANALYTICAL CHEMISTRY, 2004, 76 (15) :4446-4451
[23]   Ultrasensitive coincidence fluorescence detection of single DNA molecules [J].
Li, HT ;
Ying, LM ;
Green, JJ ;
Balasubramanian, S ;
Klenerman, D .
ANALYTICAL CHEMISTRY, 2003, 75 (07) :1664-1670
[24]   Quantum dot ligands provide new insights into erbB/HER receptor-mediated signal transduction [J].
Lidke, DS ;
Nagy, P ;
Heintzmann, R ;
Arndt-Jovin, DJ ;
Post, JN ;
Grecco, HE ;
Jares-Erijman, EA ;
Jovin, TM .
NATURE BIOTECHNOLOGY, 2004, 22 (02) :198-203
[25]   Using photon statistics to boost microscopy resolution [J].
Michalet, X ;
Weiss, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (13) :4797-4798
[26]   Quantum dots for live cells, in vivo imaging, and diagnostics [J].
Michalet, X ;
Pinaud, FF ;
Bentolila, LA ;
Tsay, JM ;
Doose, S ;
Li, JJ ;
Sundaresan, G ;
Wu, AM ;
Gambhir, SS ;
Weiss, S .
SCIENCE, 2005, 307 (5709) :538-544
[27]   Illuminating single molecules in condensed matter [J].
Moerner, WE ;
Orrit, M .
SCIENCE, 1999, 283 (5408) :1670-+
[28]  
Nelson P, 2003, BIOL PHYS
[29]   Optical detection of single molecules [J].
Nie, SM ;
Zare, RN .
ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 1997, 26 :567-596
[30]   Nanometer-localized multiple single-molecule fluorescence microscopy [J].
Qu, XH ;
Wu, D ;
Mets, L ;
Scherer, NF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (31) :11298-11303