Long‐Term Single‐Molecule Tracking in Living Cells using Weak‐Affinity Protein Labeling
2025

Long-Term Single-Molecule Tracking in Living Cells

Sample size: 160 publication 10 minutes Evidence: high

Author Information

Author(s): Claudia Catapano, Marina S. Dietz, Julian Kompa, Soohyen Jang, Petra Freund, Kai Johnsson, Mike Heilemann

Primary Institution: Johann Wolfgang Goethe-University Frankfurt

Hypothesis

Can weak-affinity protein labeling extend observation times for single-molecule tracking in living cells?

Conclusion

The study demonstrates that using exchangeable HaloTag ligands allows for long-term single-molecule tracking in living cells without significant photobleaching.

Supporting Evidence

  • The use of exchangeable HaloTag ligands allows for extended observation times of up to 30 minutes.
  • Single-molecule tracking revealed non-uniform distributions of receptor mobility.
  • Diffusion coefficients correlated with the ER nano-structure.
  • EGFR activation was monitored directly in living cells.

Takeaway

This study shows a new way to watch proteins in living cells for a long time without them disappearing from view.

Methodology

The study used exchangeable HaloTag ligands for single-particle tracking of membrane proteins in living cells.

Limitations

The study primarily focuses on specific protein interactions and may not generalize to all membrane proteins.

Statistical Information

P-Value

p<0.001

Statistical Significance

p<0.001

Digital Object Identifier (DOI)

10.1002/anie.202413117

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