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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: Jul 2, 2025

A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Targeted Single Particle Tracking with Upconverting Nanoparticles.

Oleksii Dukhno¹, Srijayee Ghosh¹, Vanille Greiner¹

¹Laboratory of Biomaging and Pathologies, UMR 7021 CNRS, University of Strasbourg, Strasbourg 67000, France.

ACS Applied Materials & Interfaces

|February 22, 2024

Summary

This summary is machine-generated.

Upconverting nanoparticles (UCNPs) offer superior performance for single particle tracking (SPT) of biomolecules in living cells. These UCNP labels overcome limitations of traditional dyes and quantum dots, enabling clearer visualization of cellular dynamics.

Keywords:

high-affinity IgE receptor nanoparticles single particle tracking single-molecule microscopy upconversion

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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

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Last Updated: Jul 2, 2025

A Protocol for Real-time 3D Single Particle Tracking

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Published on: January 3, 2018

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

Published on: October 1, 2014

Area of Science:

Biophysics
Nanotechnology
Cell Biology

Background:

Single particle tracking (SPT) visualizes biomolecule movement in real-time.
Existing labels like fluorescent dyes and quantum dots have limitations (photostability, blinking).
Upconverting nanoparticles (UCNPs) show promise as advanced SPT labels.

Purpose of the Study:

Demonstrate targeted SPT using UCNPs.
Synthesize and characterize UCNPs for cell tracking.
Compare UCNP SPT performance against conventional labels.

Main Methods:

Synthesized 30 nm doped UCNPs, coated with PEGylated polymers.
Functionalized UCNPs with immunoglobulin E (IgE) via biotin-streptavidin.
Tracked IgE receptors (FcεRI) on RBL-2H3 mast cells using UCNP-IgE labels.
Directly compared UCNP-SPT with organic dyes (AlexaFluor647) and quantum dots (QD655).

Main Results:

Developed water-dispersible, bright, and homogeneous UCNPs.
Successfully tracked FcεRI on living mast cells.
UCNP-SPT provided superior photostability and continuous luminescence.
UCNP performance was not limited by label photophysics, unlike conventional labels.

Conclusions:

UCNPs are highly effective labels for targeted SPT in live cells.
UCNP-based SPT overcomes photophysical limitations of traditional labels.
This advancement allows for more accurate study of cellular dynamics.