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Related Experiment Videos

Photon counting histograms for diffusing fluorophores.

Irina V Gopich1, Attila Szabo

  • 1Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA. gopich@nih.gov

The Journal of Physical Chemistry. B
|July 21, 2006
PubMed
Summary

This study presents photon counting histogram theory for diffusing fluorescent molecules. The developed method accurately calculates histograms, validating approximations except at low concentrations and long observation times.

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Area of Science:

  • Physical Chemistry
  • Biophysics
  • Chemical Physics

Background:

  • Photon counting histograms are crucial for analyzing fluorescent molecules.
  • Understanding molecular diffusion dynamics is essential in various scientific fields.
  • Existing methods like Fluorescence Intensity Multiple Distribution Analysis (FIMDA) have limitations.

Purpose of the Study:

  • To develop a theoretical framework for photon counting histograms of diffusing fluorescent molecules.
  • To derive analytic expressions for factorial cumulants of photon counts.
  • To validate the accuracy of the Fluorescence Intensity Multiple Distribution Analysis (FIMDA) approximation.

Main Methods:

  • Development of a theoretical model for photon counting histograms.
  • Derivation of analytic expressions for factorial cumulants.

Related Experiment Videos

  • Solving a reaction-diffusion equation for exact histogram calculation.
  • Comparison of theoretical results with numerical solutions.

    • Main Results:

    • Analytic expressions for factorial cumulants of photon counts were obtained.
    • Exact histograms can be derived by solving a reaction-diffusion equation.
    • The developed formalism correctly simplifies for immobile molecules.
    • FIMDA approximation shows high accuracy across most parameters.

    Conclusions:

    • The presented theory provides an exact method for calculating photon counting histograms.
    • FIMDA is a reliable approximation for analyzing fluorescent molecule diffusion.
    • Limitations of FIMDA are identified at low molecular concentrations and extended counting times.