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Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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...

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Related Experiment Video

Updated: Jun 3, 2026

Two-Photon-Based Photoactivation in Live Zebrafish Embryos
09:10

Two-Photon-Based Photoactivation in Live Zebrafish Embryos

Published on: December 24, 2010

Patterned two-photon photoactivation illuminates spatial reorganization in live cells.

Adam W Smith1, Alexander A Smoligovets, Jay T Groves

  • 1Howard Hughes Medical Institute, Department of Chemistry, University of California, Berkeley, California 94720, United States.

The Journal of Physical Chemistry. A
|March 12, 2011
PubMed
Summary
This summary is machine-generated.

Singular value decomposition (SVD) combined with patterned photoactivation tracks molecular movement in live cells. This method reveals dynamic actin cytoskeleton behavior in T-cells and stable actin in Cos-7 cells.

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Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist
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Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist

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Related Experiment Videos

Last Updated: Jun 3, 2026

Two-Photon-Based Photoactivation in Live Zebrafish Embryos
09:10

Two-Photon-Based Photoactivation in Live Zebrafish Embryos

Published on: December 24, 2010

In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin
05:45

In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin

Published on: July 11, 2019

Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist
07:48

Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist

Published on: April 25, 2018

Area of Science:

  • Cell biology
  • Biophysics
  • Molecular imaging

Background:

  • Photoactivatable fluorescent proteins (like PaGFP) enable precise molecular tracking.
  • Quantifying spatial reorganization of photoactivated molecules remains challenging.
  • Actin cytoskeleton dynamics are crucial for cellular processes, including immune responses.

Purpose of the Study:

  • To develop and apply a novel method for quantifying spatial redistribution of photoactivated molecules.
  • To analyze live-cell actin cytoskeleton dynamics using patterned photoactivation and SVD.
  • To compare actin behavior in motile T-cells versus more stable Cos-7 cells.

Main Methods:

  • Utilized photoactivatable green fluorescent protein (PaGFP) fused to utrophin (UtrCH) for actin labeling.
  • Employed patterned photoactivation with galvanometric scanning mirrors to write circular patterns.
  • Applied singular value decomposition (SVD) to analyze time-series radial distribution profiles of fluorophore redistribution.

Main Results:

  • SVD effectively described the time-evolution of radial distribution profiles using the first two component states.
  • In primary murine T-cells, actin filaments exhibited high mobility with inward transport (1-2s timescale), indicative of retrograde cycling.
  • In Cos-7 cells, the actin cytoskeleton was relatively stationary, showing minimal centripetal flow.

Conclusions:

  • The combination of patterned photoactivation and SVD analysis provides a powerful tool for measuring spatial redistribution dynamics in live cells.
  • This technique elucidates distinct actin cytoskeleton behaviors in different cell types, correlating with their known functions.
  • The findings offer new insights into the rapid dynamics of actin remodeling in immune cells.