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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jun 25;93(13):6264–6268. doi: 10.1073/pnas.93.13.6264

Probing the interaction between two single molecules: fluorescence resonance energy transfer between a single donor and a single acceptor.

T Ha 1, T Enderle 1, D F Ogletree 1, D S Chemla 1, P R Selvin 1, S Weiss 1
PMCID: PMC39010  PMID: 8692803

Abstract

We extend the sensitivity of fluorescence resonance energy transfer (FRET) to the single molecule level by measuring energy transfer between a single donor fluorophore and a single acceptor fluorophore. Near-field scanning optical microscopy (NSOM) is used to obtain simultaneous dual color images and emission spectra from donor and acceptor fluorophores linked by a short DNA molecule. Photodestruction dynamics of the donor or acceptor are used to determine the presence and efficiency of energy transfer. The classical equations used to measure energy transfer on ensembles of fluorophores are modified for single-molecule measurements. In contrast to ensemble measurements, dynamic events on a molecular scale are observable in single pair FRET measurements because they are not canceled out by random averaging. Monitoring conformational changes, such as rotations and distance changes on a nanometer scale, within single biological macromolecules, may be possible with single pair FRET.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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