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Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation
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Some tips and tricks for a Correlative Light Electron Microscopy workflow using stable expression of fluorescent
1BioMediTech, Faculty of Medicine and Health Technology, Tampere University, Tampere, Finland.
Methods in Cell Biology
|May 5, 2024
Summary
We present a simple Correlative Light Electron Microscopy (CLEM) workflow for 2D cell cultures. This method combines light microscopy and transmission electron microscopy (TEM) for precise molecular localization in cells.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- Correlative Light Electron Microscopy (CLEM) integrates light and electron microscopy for enhanced biological insights.
- Various CLEM approaches exist, differing in complexity and technical demands.
Purpose of the Study:
- To describe a simple and effective CLEM workflow for 2D cell cultures.
- To detail sample preparation steps for resin-embedded samples for transmission electron microscopy (TEM).
Main Methods:
- Utilizing gridded finder imaging dishes for combined light and electron microscopy.
- Employing light microscopy for initial fluorescence localization.
- Using transmission electron microscopy (TEM) for ultrastructural reference and precise localization.
Main Results:
- Demonstrated a straightforward CLEM protocol for fluorescently tagged molecules in 2D cell cultures.
- Successfully correlated light microscopy fluorescence data with TEM ultrastructure.
Conclusions:
- The presented CLEM workflow offers a powerful and accessible method for high-resolution imaging.
- This approach simplifies the process of determining the precise localization of fluorescently labeled molecules within cells.
