DOI: 10.1146/annurev.physchem.52.1.233. R<scp>ATIOMETRIC</scp>S<scp>INGLE</scp>-M<scp>OLECULE</scp>S<scp>TUDIES OF</scp>F<scp>REELY</scp>D<scp>IFFUSING</scp>B<scp>IOMOLECULES</scp>

RATIOMETRICSINGLE-MOLECULESTUDIES OFFREELYDIFFUSINGBIOMOLECULES

10.1146/annurev.physchem.52.1.233

Crossref journal-article

Annual Reviews

Annual Review of Physical Chemistry (22)

Abstract

▪ Abstract We outline recent developments in biological single-molecule fluorescence detection with particular emphasis on observations by ratiometric fluorescence resonance energy transfer (FRET) of biomolecules freely diffusing in solution. Single-molecule-diffusion methodologies were developed to minimize perturbations introduced by interactions between molecules and surfaces. Confocal microscopy is used in combination with sensitive detectors to observe bursts of photons from fluorescently labeled biomolecules as they diffuse through the focal volume. These bursts are analyzed to extract ratiometric observables such as FRET efficiency and polarization anisotropy. We describe the development of single-molecule FRET methodology and its application to the observation of the Förster distance dependence and the study of protein folding and polymer physics problems. Finally, we discuss future advances in data acquisition and analysis techniques that can provide a more complete picture of the accessible molecular information.

Bibliography

Deniz, A. A., Laurence, T. A., Dahan, M., Chemla, D. S., Schultz, P. G., & Weiss, S. (2001). RATIOMETRICSINGLE-MOLECULESTUDIES OFFREELYDIFFUSINGBIOMOLECULES. Annual Review of Physical Chemistry, 52(1), 233â253.

Authors 6

Ashok A Deniz (first)
Ted A Laurence (additional)
Maxime Dahan (additional)
Daniel S Chemla (additional)
Peter G Schultz (additional)
Shimon Weiss (additional)

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Dates

Type	When
Created	23 years ago (July 27, 2002, 7:40 a.m.)
Deposited	2 years, 4 months ago (April 23, 2023, 2:39 p.m.)
Indexed	3 weeks ago (Aug. 3, 2025, 6:53 p.m.)
Issued	23 years, 10 months ago (Oct. 1, 2001)
Published	23 years, 10 months ago (Oct. 1, 2001)
Published Print	23 years, 10 months ago (Oct. 1, 2001)

Funders 0

None

BibTeX

@article{Deniz_2001, title={R<scp>ATIOMETRIC</scp>S<scp>INGLE</scp>-M<scp>OLECULE</scp>S<scp>TUDIES OF</scp>F<scp>REELY</scp>D<scp>IFFUSING</scp>B<scp>IOMOLECULES</scp>}, volume={52}, ISSN={1545-1593}, url={http://dx.doi.org/10.1146/annurev.physchem.52.1.233}, DOI={10.1146/annurev.physchem.52.1.233}, number={1}, journal={Annual Review of Physical Chemistry}, publisher={Annual Reviews}, author={Deniz, Ashok A and Laurence, Ted A and Dahan, Maxime and Chemla, Daniel S and Schultz, Peter G and Weiss, Shimon}, year={2001}, month=oct, pages={233–253} }

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