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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Imaging Membrane Repair in Single Cells Using Correlative Light and Electron Microscopy.
Coralie Croissant1, Flora Bouvet1, Sisareuth Tan1
1Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, Pessac, France.
Current Protocols in Cell Biology
|August 8, 2018
Summary
This study presents a new protocol combining laser damage, nanoparticle staining, and advanced microscopy to visualize the molecular machinery cells use for plasma membrane repair. This method aids in understanding diseases linked to membrane repair, like muscular dystrophies and cancer metastasis.
Area of Science:
- Cell biology
- Biophysics
- Molecular medicine
Background:
- Cells can repair plasma membrane disruptions under normal conditions.
- Defective membrane repair is linked to muscular dystrophies and cancer metastasis.
- Understanding the molecular mechanisms of membrane repair is crucial for disease insights.
Purpose of the Study:
- To present a novel protocol for characterizing the molecular machinery involved in plasma membrane repair.
- To enable high-resolution imaging of membrane repair sites and protein localization.
Main Methods:
- Utilizes laser technology to induce controlled plasma membrane damage.
- Combines immunostaining with gold nanoparticles for enhanced imaging.
- Employs fluorescence microscopy for subcellular protein localization.
- Uses transmission electron microscopy (TEM) for high-resolution ultrastructural analysis of repair sites.
Main Results:
- The protocol allows detailed characterization of the molecular machinery in membrane repair.
- Fluorescence microscopy identifies protein localization in damaged cells.
- TEM provides ultrastructural insights into the repair mechanism at the disruption site.
Conclusions:
- The described protocol offers a powerful approach to study cellular membrane repair mechanisms.
- This method is applicable to human skeletal muscle cells and other cell types.
- It facilitates research into diseases associated with impaired or enhanced membrane repair.
Keywords:
correlative light and electron microscopygold nanoparticleshuman skeletal musclelasermembrane repairMore Related Videos
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