Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Different dimerization affinity and orientation of fluorescent proteins eGFP and eYFP.

Physical chemistry chemical physics : PCCP·2026
Same author

Kinetic Insights into Photoinduced Monomer-Dimer Conversion and Activation of Orange Carotenoid Protein.

The journal of physical chemistry. B·2026
Same author

The Effects of Hyperbaric Oxygen Therapy in Postoperative Patients with Ulcerative Colitis: A Retrospective Study.

The Kurume medical journal·2026
Same author

In-cell single-molecule FRET measurement of cytosolic RAF proteins to investigate the structural states and kinetics among them.

Frontiers in molecular biosciences·2025
Same author

Reaction and interaction dynamics of azobenzene-tethered DNA with T7 RNA polymerase.

Physical chemistry chemical physics : PCCP·2025
Same author

Decomposition of friction coefficients to analyze hydration effects on a C60(OH)n.

The Journal of chemical physics·2025

Related Experiment Video

Updated: Jul 5, 2026

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

Distribution analysis for single molecule FRET measurement.

Kenji Okamoto1, Masahide Terazima

  • 1Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, Kyoto 606-8502, Japan.

The Journal of Physical Chemistry. B
|May 22, 2008
PubMed
Summary

A novel numerical analysis method enhances single-pair fluorescence resonance energy transfer (sp-FRET) data analysis by using cumulative distribution functions. This approach offers superior component resolution compared to traditional histogram-based methods.

More Related Videos

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox
07:12

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Published on: July 5, 2021

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Related Experiment Videos

Last Updated: Jul 5, 2026

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox
07:12

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Published on: July 5, 2021

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Area of Science:

  • Biophysics
  • Computational Biology
  • Biochemistry

Background:

  • Single-pair fluorescence resonance energy transfer (sp-FRET) is a powerful technique for measuring distances in biological molecules.
  • Traditional analysis methods often rely on histogram plots and Gaussian fitting, which can be limited by bin size and resolution.
  • Accurate analysis of sp-FRET data is crucial for understanding molecular dynamics and interactions.

Purpose of the Study:

  • To introduce a new, bin-size-independent numerical analysis method for sp-FRET data.
  • To demonstrate the improved accuracy and efficiency of this method compared to existing techniques.
  • To validate the method's performance on simulated and experimental sp-FRET data.

Main Methods:

  • Developed a novel analysis approach plotting each data point as a cumulative distribution function.
  • Performed numerical simulations to compare the new method with Gaussian fitting on histogram plots.
  • Applied the method to experimental sp-FRET data from labeled double-strand DNA.

Main Results:

  • The proposed cumulative distribution function method showed more efficient and accurate component resolvability than Gaussian fitting.
  • Simulations indicated reliable fitting error analysis for the new method.
  • Experimental sp-FRET data from DNA revealed successful resolution of up to four subpopulations in mixtures of up to three species.

Conclusions:

  • The new numerical analysis method provides a robust and accurate approach for sp-FRET data.
  • This method overcomes limitations of traditional bin-dependent analyses, enabling better characterization of complex molecular systems.
  • The technique is effective for analyzing experimental sp-FRET data, including complex DNA structures.