Probing Single Molecule Dynamics

Xie, Xiaohui; Dunn, Robert C.

doi:10.1126/science.265.5170.361

Cited by 583 publications

(460 citation statements)

References 33 publications

(9 reference statements)

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“…The digital photobleaching is characteristic of a single molecule. The molecule in Figure 6C exhibits blinking behavior similar to that observed in previous single-molecule experiments [31][32][33] . Blinking is observed for ~15% of the molecules on the time scale of the measurement (100 ms), and this degree of blinking is less severe than for many other single-molecule emitters.…”

Section: Single-molecule Measurementssupporting

confidence: 66%

Novel Fluorophores for Single-Molecule Imaging

Willets

Ostroverkhova

et al. 2003

J. Am. Chem. Soc.

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A new class of fluorophores has been identified that can be imaged at the single-molecule level and offer additional beneficial properties such as a significant ground state dipole moment, moderate hyperpolarizability, and sensitivity to local rigidity. These molecules contain an amine donor and a dicyanodihydrofuran (DCDHF) acceptor linked by a conjugated unit (benzene, thiophene, alkene, styrene, etc.) and were originally designed to deliver both high polarizability anisotropy and dipole moment as nonlinear optical chromophores for photorefractive applications. Surprisingly, we have found that these molecules are also well-suited for single-molecule fluorescence imaging in polymers and other reasonably rigid environments. We report the bulk (ensemble) and single-molecule photophysical properties measured for six dyes in this new class of single-molecule reporters, with absorption maxima ranging from 486 to 614 nm.

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Section: Single-molecule Measurementssupporting

confidence: 66%

Novel Fluorophores for Single-Molecule Imaging

Willets

Ostroverkhova

et al. 2003

J. Am. Chem. Soc.

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“…[2][3][4][5][6]8,9 Molecular interactions and chemical dynamics can now be probed in specific local environments. There are two distinctly different time scales on which one can probe single-molecule dynamics.…”

Section: Introductionmentioning

confidence: 99%

“…Recent advances in near-field [1][2][3][4][5][6] and far-field 7-9 fluorescence microscopies have made it possible to image the emission from single molecules with high spatial resolutions and to extend the single-molecule spectroscopy work done at cryogenic temperatures 10 to the room temperature regime. [2][3][4][5][6]8,9 Molecular interactions and chemical dynamics can now be probed in specific local environments.…”

Section: Introductionmentioning

confidence: 99%

Single-Molecule Kinetics of Interfacial Electron Transfer

1997

Self Cite

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Measurements of single-molecule chemical reaction kinetics are demonstrated for interfacial electron transfer from excited cresyl violet molecules to the conduction band of indium tin oxide (ITO) or energetically accessible surface electronic states under ambient conditions by using a far-field fluorescence microscope. In this system, each single molecule exhibits a single-exponential electron transfer kinetics. A wide distribution of sitespecific electron transfer rates is observed for many single cresyl violet molecules examined. This work reveals that the physical origin of multiexponential kinetics of electron transfer in this system is the inhomogeneity of molecular interactions on the semiconductor surface of ITO. We illustrate that the singlemolecule experiments provide detailed information not obtainable from experiments conducted on large ensembles of molecules. Single-molecule kinetics is particularly useful in understanding multiexponential behavior of chemical reactions in heterogeneous systems.

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“…These include e.g., a modified version of flow cytometry [63,64 ] used for single fluorophore detection and nearfield microscopy [65][66][67], evanescent-field fluorescence microscopy (EFFM, also called total internal reflection microscopy [68]) [69], and versions of epi-fluorescence microscopy [70][71][72][73] used for single fluorophore imaging. In the case of low temperature experiments, reviewed in Ref.…”

Section: Introductionmentioning

confidence: 99%

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions

Zheng¹,

Goldner²,

Leuba³

2007

Methods

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Many technical improvements in fluorescence microscopy over the years have focused on decreasing background and increasing the signal to noise ratio (SNR). The scanning confocal fluorescence microscope (SCFM) represented a major improvement in these efforts. The SCFM acquires signal from a thin layer of a thick sample, rejecting light whose origin is not in the focal plane thereby dramatically decreasing the background signal. A second major innovation was the advent of high quantum-yield, low noise, single-photon counting detectors. The superior background rejection of SCFM combined with low-noise, high-yield detectors makes it possible to detect the fluorescence from single dye molecules. By labeling a DNA molecule or a DNA/protein complex with a donor/ acceptor dye pair, fluorescence resonance energy transfer (FRET) can be used to track conformational changes in the molecule/complex itself, on a single molecule/complex basis. In this methods paper, we describe the core concepts of SCFM in the context of a study that uses FRET to reveal conformational fluctuations in individual Holliday junction DNA molecules and nucleosomal particles. We also discuss data processing methods for SCFM.

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Probing Single Molecule Dynamics

Cited by 583 publications

References 33 publications

Novel Fluorophores for Single-Molecule Imaging

Novel Fluorophores for Single-Molecule Imaging

Single-Molecule Kinetics of Interfacial Electron Transfer

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions

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