Förster resonance energy transfer between identical chromophores (homo-FRET) has been difficult to analyze since neither emission intensity nor lifetime changes with the occurrence of homo-FRET. Herein we used a DNA scaffold to analyze homo-FRET between pyrene moieties. The DNA scaffold was modified with two pyrenes and a quencher, anthraquinone. Homo-FRET was detected by monitoring quenching of pyrene emission and the decrease in the fluorescence lifetime of pyrene. Homo-FRET efficiencies could be calculated by excluding effects of hetero-FRET. The experimentally determined efficiencies showed an excellent agreement with Förster theory. These results will inform design of novel molecular probes and light-harvesting antennae.
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