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Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy
Open accessAbstract
A recently developed, real-time spectroscopic technique, burst-integrated fluorescence lifetime (BIFL), is shown to be well suited for monitoring the individual molecular conformational dynamics of a single molecule diffusing through the microscopic, open measurement volume (≈10 fl) of a confocal epi-illuminated set-up. In a highly diluted aqueous solution of 20-mer oligonucleotide strand of DNA duplex labeled with the environment-sensitive fluorescent dye tetramethylrhodamine (TMR), fluorescence bursts indicating traces of individual molecules are registered and further subjected to selective burst analysis. The two-dimensional BIFL data allow the identification and detection of different temporally resolved conformational states. A complementary autocorrelation analysis was performed on the time-dependent fluctuations in fluorescence lifetime and intensity. The consistent results strongly support the hypothesized three-state model of the conformational dynamics of the TMR–DNA duplex with a polar, a nonpolar, and a quenching environment of TMR.
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Cited by
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Fernandes, P. B. (1998) · Current Opinion in Chemical Biology · doi:10.1016/s1367-5931(98)80089-6 · Cited by 170
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Metadata
- Authors
- Eggeling, C., Fries, J. R., Brand, L., Günther, R., & Seidel, C. A. M.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1998-02-17
- Volume
- 95
- Issue
- 4
- Pages
- 1556-1561
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1977420895
- Cited by
- 341
- References
- 0
- FWCI
- 19.02
- Type
- journal article
- Language
- en
- Open access
- green
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- Eggeling, C., Fries, J. R., Brand, L., Günther, R., & Seidel, C. A. M. (1998). Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.95.4.1556
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