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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Efficient fluorescence recovery using antifade reagents in correlative light and electron microscopy
Kiminori Toyooka1, Naeko Shinozaki-Narikawa1
1Mass Spectrometry and Microscopy Unit, RIKEN Center for Sustainable Resource Science, Suehiro-cho 1-7-22, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
Microscopy (Oxford, England)
|August 16, 2019
Summary
We developed a simple method to recover fluorescence in resin-embedded samples for correlative light and electron microscopy (CLEM). This technique enhances green fluorescent protein (GFP) signals, improving ultrastructural analysis in microscopy.
Area of Science:
- Biophysics
- Cell Biology
- Microscopy
Background:
- Correlative light and electron microscopy (CLEM) combines fluorescence and electron microscopy for ultrastructural analysis.
- Fluorescence decay in resin-embedded samples poses a significant challenge for CLEM.
- Effective detection of fluorescence is crucial for high-resolution biological imaging.
Purpose of the Study:
- To develop a method for fluorescence recovery in resin-embedded samples for CLEM.
- To enhance the detection of green fluorescent protein (GFP) signals in microscopy.
- To overcome fluorescence decay issues in sample preparation for CLEM.
Main Methods:
- Developed a fluorescence recovery method using commercially available antifade reagents.
- Applied the method to resin-embedded semi-thin sections.
- Utilized field-emission scanning electron microscopy for imaging.
Main Results:
- Successfully recovered fluorescence of GFP in resin-embedded semi-thin sections.
- Obtained CLEM images with moderately enhanced GFP signals.
- Demonstrated the efficacy of the simple fluorescence recovery method.
Conclusions:
- The developed method effectively restores fluorescence in resin-embedded samples.
- This technique simplifies and improves the quality of CLEM imaging.
- The approach offers a practical solution for enhancing ultrastructural analysis in biological research.
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