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Protein Dynamics in Living Cells

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 16, 2026

Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations

Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations

Published on: September 20, 2019

Imaging protein dynamics in live mitotic cells.

Nick P Ferenz¹, Nan Ma, Wei-Lih Lee

¹Department of Biology, University of Massachusetts, Amherst, MA 01003, USA.

Methods (San Diego, Calif.)

|January 21, 2010

Summary

This summary is machine-generated.

Researchers developed a new method using photoactivatable GFP (PA-GFP) to track individual proteins within the mitotic spindle. This technique allows detailed analysis of protein dynamics during cell division, advancing our understanding of spindle assembly and function.

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Last Updated: Jun 16, 2026

Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations

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Published on: September 20, 2019

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Live Imaging of Mitosis in the Developing Mouse Embryonic Cortex

Live Imaging of Mitosis in the Developing Mouse Embryonic Cortex

Published on: June 4, 2014

Area of Science:

Cell Biology
Molecular Biology
Biochemistry

Background:

Accurate segregation of genetic material during mitosis relies on the precise assembly and function of the mitotic spindle.
Previous studies using conventional fluorescent tags provided insights but limited detailed analysis of individual protein dynamics due to uniform fluorescence.
Understanding the dynamic behavior of mitotic spindle components is crucial for a complete picture of spindle assembly.

Purpose of the Study:

To develop and validate a method for tracking the dynamic behavior of specific protein subsets within the mitotic spindle.
To overcome limitations of conventional fluorescent tagging by utilizing photoactivatable variants of green fluorescent protein (PA-GFP).
To demonstrate the utility of PA-GFP tagging for examining protein dynamics in live mammalian and yeast cells.

Main Methods:

Development of methods to tag cellular proteins with PA-GFP.
Expression of PA-GFP-tagged proteins in live mammalian and yeast cells.
Local photoactivation of PA-GFP-tagged proteins and subsequent recording of their dynamic behavior.

Main Results:

Successful tagging and expression of proteins with PA-GFP in diverse cell types.
Demonstration of localized photoactivation and tracking of specific PA-GFP-tagged molecules.
Illustration of the method's power in analyzing the dynamics of spindle formation and function.

Conclusions:

PA-GFP tagging provides a powerful approach to study the dynamics of individual protein components within the mitotic spindle.
This method enables detailed analysis of protein behavior in live cells, overcoming limitations of traditional fluorescent probes.
The described techniques are applicable across various cell types, facilitating broader research into cell division mechanisms.