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A Protocol for Real-time 3D Single Particle Tracking
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Real-time analysis of quantum dot labeled single porcine epidemic diarrhea virus moving along the microtubules using

Wei Hou1, Yangyang Li1, Wenjie Kang1

  • 1Joint International Research Laboratory of Animal Health and Food Safety, Single Molecule Nanometry Laboratory (Sinmolab) & Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing, 210095, China.

Scientific Reports
|February 6, 2019
PubMed

Insights

Porcine epidemic diarrhea virus (PEDV) transport in live Vero cells was visualized using quantum dots and single particle tracking. PEDV exhibited distinct movement patterns along microtubules, crucial for understanding infection mechanisms.

Area of Science:

  • Virology
  • Cell Biology
  • Biophysics

Background:

  • Porcine epidemic diarrhea virus (PEDV) causes a highly contagious enteric disease in pigs.
  • Understanding PEDV infection mechanisms is crucial for disease control.
  • Previous studies lacked dynamic visualization of PEDV intracellular transport.

Purpose of the Study:

  • To visualize and characterize the dynamic transport behaviors of PEDV in live Vero cells.
  • To uncover the movement patterns of PEDV along microtubules.
  • To provide insights into the early stages of PEDV infection.

Main Methods:

  • Quantum dot labeling of PEDV to maintain infectivity.
  • Single particle tracking technique for dynamic visualization.
  • Observation of PEDV movement in live Vero cells.

Main Results:

  • PEDV displayed restricted motion with stable speed near the cell membrane.
  • PEDV exhibited a slow-fast-slow velocity pattern in the cytoplasm and near the microtubule organizing center.
  • Return movements of a small number of PEDVs were observed.

Conclusions:

  • This study provides the first dynamic characterization of PEDV transport along microtubules.
  • The findings reveal distinct intracellular movement patterns of PEDV.
  • The research offers critical references for further studies on PEDV infection mechanisms.

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