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Time-resolved burst variance analysis.

Ivan Terterov1, Daniel Nettels2, Dmitrii E Makarov3

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Summary
This summary is machine-generated.

This study introduces time-resolved burst variance analysis for quantifying biomolecular dynamics using single-molecule Förster resonance energy transfer (smFRET). This method efficiently extracts kinetic rates from diffusing molecules, requiring fewer photons and reducing measurement time.

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Area of Science:

  • Biophysics
  • Spectroscopy
  • Biomolecular Dynamics

Background:

  • Single-molecule fluorescence spectroscopy is crucial for quantifying biomolecular dynamics.
  • Single-molecule Förster resonance energy transfer (smFRET) extracts kinetic information from photon streams of donor and acceptor fluorophores.

Purpose of the Study:

  • To introduce a time-resolved burst variance analysis method.
  • To quantify kinetic rates on microsecond to millisecond timescales in smFRET experiments of diffusing molecules.

Main Methods:

  • Photon bursts are segmented, and FRET variance is calculated and compared to shot noise.
  • Segment size is systematically varied to capture dynamics across different timescales.
  • A theoretical framework is provided to extract kinetic rates from FRET variance decay.

Main Results:

  • The method quantifies kinetic rates for diffusing molecules in smFRET experiments.
  • It captures dynamics across microsecond to millisecond timescales.
  • The approach requires fewer photons compared to alternative methods like FCS and recurrence analysis.

Conclusions:

  • Time-resolved burst variance analysis offers an efficient way to study biomolecular dynamics.
  • Reduced photon requirements lead to shorter measurement times.
  • This method enhances the utility of smFRET for kinetic studies.