Imaging and Time-Resolved Spectroscopy of Single Molecules at an Interface

Macklin, John J.; Trautman, J. K.; Harris, T. D.; Brus, L. E.

doi:10.1126/science.272.5259.255

Cited by 405 publications

(405 citation statements)

References 20 publications

Supporting

Mentioning

381

Contrasting

Unclassified

Order By: Relevance

“…The experiments reported here were performed on very dilute samples using a conventional inverted fluorescence microscope (Nikon, Diaphot 300), similar to that discussed in other recent reports. 8,9 The output of a mode-locked YAG:Nd laser (Coherent Antares) is frequency doubled to 532 nm (2 W, 80 ps fwhm, 76 MHz) and used to synchronously pump a cavity-dumped dye laser (Coherent 702) operating with Rhodamine 6G lasing at 570 nm. The output of the dye laser (10 ps fwhm pulses train at 8 MHz repetition rate) was passed through a prism to eliminate spontaneous emission and was intensity stabilized by a laser stabilization accessory (Liconix 50SA).…”

Section: Methodsmentioning

confidence: 99%

“…On the picosecond to nanosecond time scale, excited state processes, such as radiative and nonradiative energy transfer processes, have been studied by repetitive excitation with short laser pulses. [4][5][6]8 In this Letter, we demonstrate an example for measuring chemical kinetics on this fast time scale.…”

Section: Introductionmentioning

confidence: 93%

“…[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%

See 2 more Smart Citations

Single-Molecule Kinetics of Interfacial Electron Transfer

1997

View full text Add to dashboard Cite

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.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 93%

See 1 more Smart Citation

Single-Molecule Kinetics of Interfacial Electron Transfer

1997

View full text Add to dashboard Cite

show abstract

“…Using such devices, one may obtain the images of single molecules-as was demonstrated on the green fluorescent protein (GFP) of Aequora victoria [130]. In recent years, the possibility of studying nanoobjects with the aid of SNOM has arisen [108] -along with far-field microscopy [131]. Wide-field microscopy with total internal reflection enabled, for instance, to observe individual ATP turnovers by a single myosin molecule [132] and, also, to observe diffusion of partly immobilized molecules into the lipid membrane at video rate.…”

Section: Potentialities Of Confocal and Near-field Microscopiesmentioning

confidence: 99%

Nanotechnologies in proteomics

Ivanov

Govorun

Быков³

et al. 2006

Proteomics

View full text Add to dashboard Cite

Progress in proteomic researches is largely determined by development and implementation of new methods for the revelation and identification of proteins in biological material in a wide concentration range (from 10 23 M to single molecules). The most perspective approaches to address this problem involve (i) nanotechnological physicochemical procedures for the separation of multicomponent protein mixtures; among these of particular interest are biospecific nanotechnological procedures for selection of proteins from multicomponent protein mixtures with their subsequent concentration on solid support; (ii) identification and counting of single molecules by use of molecular detectors. The prototypes of biospecific nanotechnological procedures, based on the capture of ligand biomolecules by biomolecules of immobilized ligate and the concentration of the captured ligands on appropriate surfaces, are well known; these are affinity chromatography, magnetic biobeads technology, different biosensor methods, etc. Here, we review the most promising nanotechnological approaches for selection of proteins and kinetic characterization of their complexes based on these biospecific methods with subsequent MS/MS identification of proteins and protein complexes. Two major groups of methods for the analysis and identification of individual molecules and their complexes by use of molecular detectors will be reviewed: scanning probe microscopy (SPM) (including atomic-force microscopy) and cryomassdetector technology.

show abstract

“…The adaptation of confocal microscopy [78] first to detect [79,80] and then to image [81,82] single fluorophores at room temperature made the technique widely available. The first demonstration of spFRET from immobilized molecules at room temperature was performed using a near-field microscope [83].…”

Section: Introductionmentioning

confidence: 99%

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

Zheng¹,

Goldner²,

Leuba³

2007

Methods

View full text Add to dashboard Cite

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.

show abstract

Imaging and Time-Resolved Spectroscopy of Single Molecules at an Interface

Cited by 405 publications

References 20 publications

Single-Molecule Kinetics of Interfacial Electron Transfer

Single-Molecule Kinetics of Interfacial Electron Transfer

Nanotechnologies in proteomics

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

Contact Info

Product

Resources

About