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Related Concept Videos

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 7, 2026

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
11:28

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

Published on: October 1, 2014

Time-resolved three-dimensional molecular tracking in live cells.

Nathan P Wells1, Guillaume A Lessard, Peter M Goodwin

  • 1Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, United States.

Nano Letters
|October 21, 2010
PubMed
Summary
This summary is machine-generated.

We developed a new method to track quantum dot (QD) labeled proteins in live cells, enabling detailed 3D motion analysis and spectroscopy. This technique offers enhanced tracking range and lower power, revealing nanoscale cell surface features and endocytosis dynamics.

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Last Updated: Jun 7, 2026

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Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix
10:26

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Published on: December 22, 2011

Area of Science:

  • Biophysics
  • Cell Biology
  • Quantum Dot Technology

Background:

  • Tracking individual molecules in live cells is crucial for understanding cellular processes.
  • Existing 3D tracking methods have limitations in range, excitation power, and spectroscopic capabilities.

Purpose of the Study:

  • To present a novel method for high-resolution 3D tracking of individual quantum dot (QD) labeled proteins in live cells.
  • To demonstrate the advantages of this method over existing techniques, including enhanced tracking range and lower excitation power.
  • To showcase the application of this method in studying cellular dynamics, such as receptor trafficking and endocytosis.

Main Methods:

  • Utilizes four overlapping confocal volume elements with active feedback for 5 ms tracking.
  • Employs quantum dots (QDs) for labeling proteins, enabling 3D molecular motion analysis.
  • Incorporates time-resolved spectroscopy, including fluorescence lifetime and correlation spectroscopy.

Main Results:

  • Achieved a 10-micrometer Z-tracking range with substantially lower excitation powers (15 microW).
  • Demonstrated fluorescence photon antibunching of individual QD-labeled proteins in live cells for the first time.
  • Successfully tracked individual dye-labeled nucleotides (Cy5-dUTP) at biologically relevant rates.
  • Revealed 3D nanoscale features of the cell surface topology by tracking IgE-FcεRI receptors.
  • Captured and tracked ligand-mediated endocytosis and rapid vesicular transit (~950 nm/s) of QD-labeled receptors.

Conclusions:

  • The developed method provides significant advantages for 3D molecular tracking in live cells.
  • Enables unprecedented insights into the spatiotemporal dynamics of cellular processes at the nanoscale.
  • Opens new avenues for studying molecular interactions and trafficking within live cells using advanced spectroscopy.