Extraction of lifetime distributions from fluorescence decays with application to DNA-base analogues

Aoife C. Fogarty, Anita C. Jones, Philip J. Camp

Research output: Contribution to journalArticlepeer-review

Abstract

Several important aspects of fluorescence decay analysis are addressed and tested against new experimental measurements. A simulated-annealing method is described for deconvoluting the instrument response function from a measured fluorescence decay to yield the true decay, which is more convenient for subsequent fitting. The method is shown to perform well against the conventional approach, which is to fit a convoluted fitting function to the experimentally measured decay. The simulated annealing approach is also successfully applied to the determination of an instrument response function using a known true fluorescence decay (for rhodamine 6G). The analysis of true fluorescence decays is considered critically, focusing specifically on how a distribution of decay constants can be incorporated in to a fit. Various fitting functions are applied to the true fluorescence decays of 2-aminopurine in water-dioxane mixtures, in a dinucleotide, and in DNA duplexes. It is shown how a suitable combination of exponential decays and non-exponential decays (based on a Gamma distribution of decay constants) can provide fits of equal quality to the conventional multi-exponential fits used in the majority of previous studies, but with fewer fitting parameters. Crucially, the new approach yields decay-constant distributions that are physically more meaningful than those corresponding to the conventional multi-exponential fit. The methods presented here should find wider application, for example to the analysis of transient-current or optical decays and in Forster resonance energy transfer (FRET).

Original languageEnglish
Pages (from-to)3819-3830
Number of pages12
JournalPhysical Chemistry Chemical Physics
Volume13
Issue number9
DOIs
Publication statusPublished - 2011

Keywords

  • TIME-RESOLVED FLUORESCENCE
  • RESONANCE ENERGY-TRANSFER
  • MAXIMUM-ENTROPY METHOD
  • ACTIVE-SITE
  • ACCEPTOR DISTRIBUTION
  • CHARGE-TRANSPORT
  • EXCITED-STATE
  • 77 K
  • 2-AMINOPURINE
  • DYNAMICS

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