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Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells
Published on: September 7, 2018
Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron
F G A Faas1, M Bárcena, A V Agronskaia
1Department of Molecular Cell Biology, Section Electron Microscopy, Leiden University Medical Center, P.O. Box 9600, 2300 RC, Leiden, The Netherlands.
Journal of Structural Biology
|December 25, 2012
Summary
Cryogenic integrated light and electron microscopy preserves native biological states for high-resolution imaging. This advanced technique enhances fluorescence signal and reduces photobleaching, improving correlative microscopy success.
Area of Science:
- Biological imaging
- Microscopy techniques
Background:
- Correlative light and electron microscopy (CLEM) integrates fluorescence and electron microscopy for multi-resolution biological studies.
- Previous integrated CLEM approaches operated at room temperature, posing challenges for hydrated specimens and fluorescence signal integrity.
Purpose of the Study:
- To present an integrated approach for correlative microscopy under cryogenic conditions.
- To highlight the advantages of cryo-CLEM over room temperature methods for biological sample analysis.
Main Methods:
- Development of an integrated light and electron microscope system operating at cryogenic temperatures.
- Application of the cryo-integrated system to various biological samples, including bacteria, microtubules, fibroblasts, and mammalian cells.
Main Results:
- Cryo-integrated CLEM preserves the native hydrated state of biological specimens.
- Maintained fluorescence signal integrity without quenching from heavy atom stains.
- Demonstrated reduced photobleaching compared to room temperature methods.
- Successfully applied to diverse biological applications, including bacteria detection and subcellular structure analysis.
Conclusions:
- Cryo-integrated light and electron microscopy offers significant advantages for high-resolution imaging of biological systems.
- This technique is crucial for preserving sample integrity and fluorescence signals, enabling more accurate morphological and structural studies.
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