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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
Published on: June 24, 2013
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Correlative light and electron microscopy: from live cell dynamic to 3D ultrastructure
Coralie Spiegelhalter1, Jocelyn F Laporte, Yannick Schwab
1Imaging Center, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France.
Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2013
Summary
Correlative light and electron microscopy (CLEM) integrates live-cell imaging with transmission electron microscopy (TEM) for ultrastructural analysis. This method precisely correlates dynamic cellular events observed with light microscopy to their ultrastructure using high-pressure freezing.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biophysics
Background:
- Correlative light and electron microscopy (CLEM) combines data from light microscopy (LM) and electron microscopy (EM).
- Existing CLEM methods vary widely in LM and EM combinations and specimen types.
- This work focuses on correlating live-cell LM with transmission EM (TEM).
Purpose of the Study:
- To present a method for correlating live-cell LM observations with TEM ultrastructure.
- To enable straightforward correlation of specific cellular structures between LM and EM.
- To capture dynamic events observed in live cells at the ultrastructural level.
Main Methods:
- Live-cell light microscopy (LM) acquisition.
- Development of a precise coordinate system for correlating LM and EM data.
- High-pressure freezing for sample fixation.
- Transmission electron microscopy (TEM) for ultrastructural analysis.
Main Results:
- A method for precise spatial correlation between LM and TEM was established.
- High-pressure freezing effectively preserved dynamic cellular events for ultrastructural study.
- Successful integration of live-cell dynamics with ultrastructural details.
Conclusions:
- The described CLEM technique allows for detailed ultrastructural analysis of dynamic events observed in live cells.
- Precise coordinate mapping is crucial for accurate LM-EM correlation.
- High-pressure freezing is a suitable fixation method for capturing transient cellular processes in CLEM.
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