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Probing the interaction between two single molecules: fluorescence resonance energy transfer between a single donor and a single acceptor.
Open accessAbstract
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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Cited by
Bout, D. A. V., Yip, W.-T., Hu, D., Fu, D.-K., Swager, T. M., & Barbara, P. F. (1997) · Science · doi:10.1126/science.277.5329.1074 · Cited by 521
Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study
Hellenkamp, B., Schmid, S., Doroshenko, O., Opanasyuk, O., Kühnemuth, R., Rezaei Adariani, S., Ambrose, B., Aznauryan, M., Barth, A., Birkedal, V., et al. (2018) · Nature Methods · doi:10.1038/s41592-018-0085-0 · Cited by 519
Self-citationSwitchable Fluorophores for Single-Molecule Localization Microscopy
Li, H., & Vaughan, J. C. (2018) · Chemical Reviews · doi:10.1021/acs.chemrev.7b00767 · Cited by 319
Real-Time Observation of RecA Filament Dynamics with Single Monomer Resolution
Joo, C., McKinney, S. A., Nakamura, M., Rasnik, I., Myong, S., & Ha, T. (2006) · Cell · doi:10.1016/j.cell.2006.06.042 · Cited by 305
Self-citationExtreme Bendability of DNA Less than 100 Base Pairs Long Revealed by Single-Molecule Cyclization
Vafabakhsh, R., & Ha, T. (2012) · Science · doi:10.1126/science.1224139 · Cited by 301
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Metadata
- Authors
- Ha, T., Enderle, T., Ogletree, D. F., Chemla, D. S., Selvin, P. R., & Weiss, S.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1996-06-25
- Volume
- 93
- Issue
- 13
- Pages
- 6264-6268
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2000057317
- Cited by
- 1,242
- References
- 0
- FWCI
- 14.80
- Type
- journal article
- Language
- en
- Open access
- green
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- Ha, T., Enderle, T., Ogletree, D. F., Chemla, D. S., Selvin, P. R., & Weiss, S. (1996). Probing the interaction between two single molecules: fluorescence resonance energy transfer between a single donor and a single acceptor. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.93.13.6264
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