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. 2019 Aug 2;24(15):2826. doi: 10.3390/molecules24152826

Figure 17.

Figure 17

Tetrahedral detection tracking of intramolecular FRET within double-stranded DNA in 90% glycerol-water solution. (A) X, Y, Z position and fluorescent intensity of AF488 (Alexa Fluor 488; ex: 490 nm; em: 525 nm)-DNA-AF594 (Alexa Fluor 4594; ex: 590 nm; em: 617 nm) molecule. (B) Fluorescence intensity and FRET efficiency as a function of time for the trajectory shown in (A). (C) Distribution of FRET efficiency of AF488-DNA molecules (control sample). (D) Distribution of FRET efficiency of AF488-DNA-AF594 molecules. Reprinted with permission from reference [71]. Copyright 2018 American Chemical Society. (E–I) Measurement of the hybridization kinetics of freely diffusing ssDNA molecules in solution by tetrahedral detection 3D tracking. (E) Scheme of the donor-quencher system for the DNA hybridization kinetics measurement. Red ball denotes ATTO633 (ex: 630 nm; em: 651 nm) reporter and grey ball denotes Iowa Black quencher. (F) Fluorescence lifetimes of ATTO633 with different donor-quencher separation distances. (G) Lifetime measurement in 3D tracking of ssDNA in 70 wt% glycerol solution. The fluorescence lifetime switching indicates transient annealing and melting events. (H) Lifetime histogram built from (G) showing two states. (I) Apparent annealing rate (kon) and melting rate (koff) as a function of quencher concentration shows the annealing rate is quencher concentration dependent. Adapted from reference [105]. Copyright 2017 the Royal Society of Chemistry.