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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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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: Aug 23, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
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Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells.

Chao Jiang1, Shuo-Xing Dou2, Peng-Ye Wang3

  • 1School of Systems Science and Institute of Nonequilibrium Systems, Beijing Normal University, Beijing 100875, China; Beijing National Laboratory for Condensed Matter Physics and Laboratory of Soft Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190, China; School of Physical Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.

STAR Protocols
|November 1, 2022
PubMed
Summary

This study presents a new protocol for 3D single-particle tracking in living cells. It enables detailed analysis of intracellular transport dynamics for various cellular components.

Keywords:
Cell BiologyMicroscopySingle-molecule Assays

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Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Intracellular transport is crucial for cellular functions.
  • Understanding molecular and vesicular movement is key to cell physiology.

Purpose of the Study:

  • To describe a protocol for 3D single-particle tracking in living cells.
  • To enable quantification of intracellular transport dynamics.

Main Methods:

  • Utilizes a two-focal imaging system.
  • Includes analytical steps for quantifying 3D transport dynamics.

Main Results:

  • The protocol allows for characterization of diffusion and trafficking.
  • Applicable to macromolecules, nanoparticles, and endocytic vesicles.

Conclusions:

  • This method provides a tool for studying intracellular transport.
  • Facilitates research on the movement of various components within adherent cells.