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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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Updated: May 5, 2026

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy
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Hour-Long, Kilohertz Sampling Rate Three-Dimensional Single-Virus Tracking in Live Cells Enabled by StayGold

Yuxin Lin1, Jack Exell1, Haoting Lin1

  • 1Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.

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|May 29, 2024
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Summary

New StayGold fluorescent proteins enable hour-long, high-speed 3D single-virus tracking (SVT) in live cells. This breakthrough allows detailed observation of viral dynamics and the entire viral life cycle.

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Area of Science:

  • Virology
  • Biophysics
  • Cell Biology

Background:

  • Viral infection dynamics occur across broad spatiotemporal scales.
  • High-speed, long-duration tracking of viruses requires exceptionally bright and photostable fluorescent labels.
  • Existing fluorescence protein variants often lack the necessary photostability and signal intensity for extended single-virus tracking (SVT).

Purpose of the Study:

  • To develop a novel labeling strategy for enhanced single-virus tracking (SVT).
  • To utilize the enhanced properties of the StayGold fluorescent protein for improved viral dynamics studies.
  • To achieve hour-long, high-temporal-resolution 3D SVT in live cells.

Main Methods:

  • Engineered virus-like particles (VLPs) fused with the StayGold fluorescent protein (StayGold-VLPs).
  • Performed 3D single-virus tracking (SVT) in live cells at a sampling rate of 1000 localizations per second (kHz).
  • Utilized StayGold's superior photostability and signal intensity for prolonged imaging.

Main Results:

  • StayGold-VLPs enabled hour-long 3D SVT, significantly extending tracking duration compared to previous methods.
  • Achieved high temporal resolution (kHz sampling rate) over extended periods, capturing detailed viral trafficking dynamics.
  • Demonstrated StayGold's enhanced photostability and signal intensity under identical illumination.

Conclusions:

  • StayGold-VLPs offer a simple, generalizable VLP labeling strategy for superior SVT performance.
  • This approach significantly increases the data obtainable from single viral trajectories.
  • Enables unprecedented long-term observation of viral dynamics, potentially including the entire viral life cycle.