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Updated: Jul 10, 2025
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Detection and Quantification of Tunneling Nanotubes Using 3D Volume View Images
Published on: August 31, 2022
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Identification and Characterization of Tunneling Nanotubes for Intercellular Trafficking
Inés Sáenz-de-Santa-María1, J Michael Henderson1, Anna Pepe1
1Unité de Trafic Membranaire et Pathogénèse, Département de Biologie Cellulaire et Infection, Institut Pasteur, CNRS UMR3691, Paris, France.
Current Protocols
|November 23, 2023
Summary
Tunneling nanotubes (TNTs) are crucial for cell communication and material transfer. This protocol provides methods to identify, characterize, and assess the function of these delicate cellular structures.
Area of Science:
- Cell Biology
- Cytoskeletal Dynamics
- Intercellular Communication
Background:
- Tunneling nanotubes (TNTs) are actin-based membrane structures facilitating direct cell-to-cell transfer.
- Their delicate nature and heterogeneity pose challenges for accurate study.
- Evidence suggests TNTs are involved in intercellular communication, pathogen spread, and protein transfer.
Purpose of the Study:
- To provide a comprehensive guideline for the identification and characterization of TNTs in cell culture.
- To offer optimized protocols for studying TNT formation, structure, and function.
- To address common pitfalls in TNT research.
Main Methods:
- Optimized protocols for TNT identification and cytoskeletal element analysis.
- Live-cell imaging to monitor TNT formation and dynamics.
- Micropatterning and cryo-electron microscopy (cryo-EM) for structural characterization.
- Assessment of TNT-mediated intercellular transfer of cellular components.
Main Results:
- Detailed methods for distinguishing TNTs from other cellular protrusions.
- Protocols for evaluating TNT frequency and lifetime.
- Techniques to confirm functional TNTs through cargo transfer assays.
- Cryo-EM enables detailed ultrastructural analysis of TNTs.
Conclusions:
- This protocol offers a robust framework for reliable TNT research.
- Standardized methods are essential for advancing our understanding of TNT roles in cell biology.
- Accurate characterization of TNTs will facilitate studies on their involvement in health and disease.
Keywords:
correlative cryo-electron microscopy (cryo-EM)cytoskeletonintercellular communicationmicropatterningtunneling nanotubes (TNTs)More Related Videos
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