Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy.
作者： C. Eggeling , J. R. Fries , L. Brand , R. Gunther , C. A. M. Seidel
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摘要: 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 microscopic, open measurement volume (≈10 fl) confocal epi-illuminated set-up. In highly diluted aqueous solution 20-mer oligonucleotide strand DNA duplex labeled with environment-sensitive fluorescent dye tetramethylrhodamine (TMR), bursts indicating traces molecules are registered and further subjected selective burst analysis. The two-dimensional BIFL data allow identification detection different temporally resolved states. complementary autocorrelation analysis was performed on time-dependent fluctuations in intensity. consistent results strongly support hypothesized three-state model TMR–DNA polar, nonpolar, quenching environment TMR.
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