To explore the dynamics of biomols., tracing the kinetics of photo-induced chem. reactions via the triplet excited state (T1) of probe mols. offers a timescale that is about 106 times wider than via the singlet excited state (S1). Using cyclooctatetraene (COT) as a triplet energy acceptor and at the same time as a photostabilizer, the triplet-triplet energy transfer (TTET) kinetics governed by oligonucleotide (oligo) dynamics were studied at the single-mol. level by measuring fluorescence blinking. TTET kinetics measurement allowed us to access the length- and sequence-dependent dynamics of oligos and realize the single-mol. detection of a model microRNA biomarker. In sharp contrast to the singlet-singlet Foerster resonance energy transfer (FRET) that occurs in the 1-10 nm range, TTET requires a Van der Waals contact. The present method is thus a complementary method to FRET and provides direct information on biomol. dynamics on the ﾎｼs to ms timescale.
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