Jove
Visualize
Contact Us

Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.0K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.0K

You might also read

Related Articles

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

Sort by
Same author

A Unified Platform for FCS and RICS Analysis with Advanced Statistical Inference.

ACS omega·2026
Same author

Lifelong creatine deficiency leads to augmented sarcoplasmic reticulum calcium release but not heart failure.

American journal of physiology. Heart and circulatory physiology·2025
Same author

Higher AMPK activation in mouse oxidative compared with glycolytic muscle does not correlate with LKB1 or CaMKKβ expression.

American journal of physiology. Endocrinology and metabolism·2024
Same author

Involvement of Escherichia coli YbeX/CorC in ribosomal metabolism.

Molecular microbiology·2024
Same author

Simple analysis of gel images with IOCBIO Gel.

BMC biology·2023
Same author

Cardiomyocytes from female compared to male mice have larger ryanodine receptor clusters and higher calcium spark frequency.

The Journal of physiology·2023
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 Experiment Video

Updated: May 11, 2025

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions
11:22

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

Published on: January 30, 2018

10.0K

Statistical analysis of fluorescence intensity transients with Bayesian methods.

Hamed Karimi1, Martin Laasmaa1, Margus Pihlak2

  • 1Laboratory of Systems Biology, Department of Cybernetics, Tallinn University of Technology, Tallinn, Estonia.

Science Advances
|April 18, 2025
PubMed
Summary

We developed fluorescence intensity trace statistical analysis (FITSA), a new method for studying molecular interactions in live cells. FITSA provides accurate results faster and with less laser light than traditional fluorescence correlation spectroscopy (FCS).

More Related Videos

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
09:30

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Published on: January 18, 2017

11.9K
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

2.4K

Related Experiment Videos

Last Updated: May 11, 2025

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions
11:22

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

Published on: January 30, 2018

10.0K
Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
09:30

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Published on: January 18, 2017

11.9K
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

2.4K

Area of Science:

  • Biophysics
  • Cell Biology
  • Quantitative Biology

Background:

  • Single-molecule analysis in live cells is crucial for understanding molecular dynamics.
  • Fluorescence Correlation Spectroscopy (FCS) is a common technique but suffers from high laser power, long acquisition times, and complex data fitting.
  • Phototoxicity and statistical assumptions limit the reliability and efficiency of conventional FCS.

Purpose of the Study:

  • To introduce Fluorescence Intensity Trace Statistical Analysis (FITSA) as a novel Bayesian method for analyzing single-molecule fluorescence data.
  • To demonstrate FITSA's advantages over traditional FCS in terms of speed, stability, and photon efficiency.
  • To reduce phototoxicity and improve the accuracy of molecular dynamics studies.

Main Methods:

  • Developed FITSA, a Bayesian approach that directly analyzes fluorescence intensity traces.
  • Compared FITSA's performance against conventional FCS using simulated and experimental data.
  • Evaluated parameter estimation accuracy, convergence speed, and photon requirements.

Main Results:

  • FITSA demonstrated faster and more stable convergence compared to existing methods.
  • FITSA achieved comparable precision to FCS with substantially fewer photons.
  • The method proved robust even with shorter measurement times than conventional FCS.
  • FITSA's accuracy improved significantly when accounting for statistical dependencies often overlooked in FCS.

Conclusions:

  • FITSA offers a more efficient and less phototoxic alternative to FCS for quantitative molecular analysis.
  • This advancement enables more reliable and faster studies of molecular interactions in live cells.
  • FITSA represents a significant improvement for single-molecule biophysics and related fields.