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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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Updated: Dec 9, 2025

4D Imaging of Protein Aggregation in Live Cells

4D Imaging of Protein Aggregation in Live Cells

Published on: April 5, 2013

AggFluor: Fluorogenic Toolbox Enables Direct Visualization of the Multi-Step Protein Aggregation Process in Live

Charles H Wolstenholme, Hang Hu, Songtao Ye

Journal of the American Chemical Society

|September 11, 2020

Summary

Researchers developed AggFluor, a novel chemical toolbox for live cell imaging of protein aggregates. This tool differentiates soluble oligomers from insoluble aggregates, aiding disease research.

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Last Updated: Dec 9, 2025

4D Imaging of Protein Aggregation in Live Cells

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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Published on: April 25, 2021

Area of Science:

Biochemistry
Cell Biology
Chemical Biology

Background:

Protein misfolding and aggregation are linked to incurable diseases.
Current methods lack the ability to differentiate aggregate conformations in live cells.

Purpose of the Study:

To present AggFluor, a chemical toolbox for live cell imaging and differentiation of protein aggregate conformations.
To enable dual-color imaging for distinguishing soluble oligomers from insoluble aggregates.

Main Methods:

Development of AggFluor, a series of molecular rotor fluorophores based on green fluorescent protein chromophores.
Utilizing differential fluorescence turn-on based on viscosity sensitivity within aggregates.
Employing a dual-color imaging strategy for live cell analysis.

Main Results:

AggFluor probes exhibit differential fluorescence in soluble oligomers versus insoluble aggregates.
A novel dual-color imaging strategy was established to distinguish these conformations.
Demonstrated the use of proteostasis regulators to modulate aggregate formation and disassembly.

Conclusions:

AggFluor represents the first rationally designed set of molecular rotor fluorophores with broad viscosity sensitivity.
This toolbox advances live cell imaging of protein aggregation.
AggFluor can be applied to study other biological processes involving local viscosity changes.