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

T. Ha, Th Enderle, D. F. Ogletree, D. S. Chemla, P. R. Selvin, S. Weiss

Research output: Contribution to journalArticle

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 he possible with single pair FRET.

Original languageEnglish (US)
Pages (from-to)6264-6268
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number13
DOIs
StatePublished - Jun 25 1996

ASJC Scopus subject areas

  • General

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