Photobleaching statistics in single-molecule on-/off-time distributions

Haw Yang, Hao Li, Tao Liu

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


The on- and and off-time distributions from fluorescence single-molecule experiments are widely used to extract kinetics parameters with the goal to provide a quantitative description for the molecule's behavior on the ensemble level. Such experiments are inevitably influenced by photobleaching, where the fluorescent probe transitions to a nonemissive state. Yet, it appears that few reports went beyond acknowledging this unavoidable complication; in fact, it has so far been ignored when evaluating off-time distributions. Here, we present a theoretical framework that allows the derivation of analytical equations in which photobleaching kinetics are rigorously incorporated. Unexpectedly, our results indicate that the off-time distribution should be nonexponential even when all the rate processes are single exponential. With the analytical theory understood and demonstrated as easy to implement, such ubiquitous photochemical processes can now be readily included in routine experimental analyses.

Original languageEnglish (US)
Article number174101
JournalJournal of Chemical Physics
Issue number17
StatePublished - Nov 7 2019

All Science Journal Classification (ASJC) codes

  • General Physics and Astronomy
  • Physical and Theoretical Chemistry


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