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Related Experiment Video

Updated: May 31, 2026

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

Measuring FRET using time-resolved FLIM.

Penny E Morton1, Maddy Parsons

  • 1Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK.

Methods in Molecular Biology (Clifton, N.J.)
|July 13, 2011
PubMed
Summary
This summary is machine-generated.

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This study details a method using Förster or fluorescent resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) to detect protein interactions and map cell migration dynamics in fixed cells.

Area of Science:

  • Cell biology
  • Biophysics
  • Microscopy

Background:

  • Cell migration relies on protein complex formation and post-translational modifications.
  • Förster or fluorescent resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) are advanced techniques for studying molecular interactions.
  • These methods allow for the spatial localization and detection of protein-protein interactions within cells.

Purpose of the Study:

  • To describe a protocol for detecting FRET between GFP- and mRFP1-tagged proteins.
  • To enable the mapping of protein activation states and the dynamics of protein complexes involved in cell movement.
  • To provide a comprehensive guide for researchers using FRET/FLIM in fixed adherent cells.

Main Methods:

  • Utilizing Förster or fluorescent resonance energy transfer (FRET) for proximity-based sensing.

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Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET
13:38

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Published on: January 27, 2012

Related Experiment Videos

Last Updated: May 31, 2026

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

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

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET
13:38

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Published on: January 27, 2012

  • Employing fluorescence lifetime imaging microscopy (FLIM) for sensitive FRET detection.
  • Detailed protocol covering sample preparation, data acquisition, and analysis for fixed adherent cells.

    • Main Results:

    • Demonstrated detection of FRET between specifically tagged proteins (GFP and mRFP1).
    • Successfully mapped protein interactions and complex dynamics relevant to cell migration.
    • Established a reliable protocol for FRET/FLIM analysis in fixed cell systems.

    Conclusions:

    • FRET/FLIM is a powerful technique for investigating protein-protein interactions in cell migration.
    • The described protocol facilitates the study of molecular mechanisms underlying cell movement.
    • This method provides insights into protein complex formation, localization, and activation states.