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

In Remembrance of Professor Zelig Eshhar: <i>A Life Committed to CAR-T</i>.

Human gene therapy·2026
Same author

Modulation of Homer1 EVH1 domain internal dynamics by putative autism-associated mutations.

FEBS letters·2026
Same author

Moving NMR infrastructures to remote access capabilities.

Progress in nuclear magnetic resonance spectroscopy·2026
Same author

In Remembrance of Professor Zelig Eshhar: <i>A Life Committed to CAR-T</i>.

Human gene therapy·2026
Same author

<i>N</i>-(ω-Guanidinoalkyl)amides of Eremomycin: New Type of Glycopeptides with High Potency against Planktonic Forms and Biofilms of Resistant Bacteria.

ACS infectious diseases·2026
Same author

A Fluorescent Protein-Tagged Scorpion Toxin with Picomolar Affinity for the Kv1.3 K <b><sup>+</sup></b> Channel.

ACS omega·2026

Related Experiment Video

Updated: Mar 23, 2026

Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques
08:28

Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques

Published on: November 2, 2018

8.9K

rFRET: A comprehensive, Matlab-based program for analyzing intensity-based ratiometric microscopic FRET experiments.

Peter Nagy1, Ágnes Szabó1,2, Tímea Váradi1

  • 1Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|March 23, 2016
PubMed
Summary

This study introduces a user-friendly Matlab application for quantitative microscopic Förster resonance energy transfer (FRET) analysis. The software enhances FRET efficiency measurements by offering advanced calibration, correction factors, and data visualization tools.

Keywords:
FRETimage analysismicroscopyshot noise

More Related Videos

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors
10:34

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

Published on: August 20, 2012

23.9K
Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
19:05

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

Published on: October 30, 2015

13.0K

Related Experiment Videos

Last Updated: Mar 23, 2026

Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques
08:28

Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques

Published on: November 2, 2018

8.9K
FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors
10:34

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

Published on: August 20, 2012

23.9K
Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
19:05

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

Published on: October 30, 2015

13.0K

Area of Science:

  • Biophysics
  • Cell Biology
  • Biochemistry

Background:

  • Förster resonance energy transfer (FRET) is crucial for studying protein interactions and conformations.
  • Current microscopic FRET analysis tools lack quantitative calibration, parameter distribution visualization, and gating capabilities, limiting accuracy.
  • Existing limitations hinder precise assessment of protein clustering and conformational changes using FRET microscopy.

Purpose of the Study:

  • To develop a quantitative, user-friendly Matlab application for microscopic FRET measurements.
  • To address the limitations of existing FRET analysis software by incorporating advanced features.
  • To improve the accuracy and reliability of FRET efficiency calculations in microscopy.

Main Methods:

  • Development of a graphical user interface (GUI)-controlled Matlab application.
  • Implementation of overspill and spectroscopic correction factor calculations.
  • Inclusion of multiple calculation modes: pixel-by-pixel, regression, maximum likelihood estimation (MLE), and summed intensities.
  • Integration of histogram and dot plot visualizations for parameter distribution analysis.
  • Capability for gating on plots and mask images to refine calculations.

Main Results:

  • The application accurately calculates FRET efficiency with necessary correction factors.
  • It provides visualization of parameter distributions through histograms and dot plots.
  • Multiple calculation methods (MLE, regression, summed intensities) offer flexibility and robustness.
  • Confidence intervals can be estimated via parameter simulations.

Conclusions:

  • The developed Matlab application significantly enhances quantitative microscopic FRET analysis.
  • It overcomes limitations of existing tools by providing robust calibration, visualization, and flexible calculation modes.
  • The software offers valuable insights into the reliability and distribution of FRET parameters, advancing protein interaction studies.