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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
Published on: June 24, 2013
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Live correlative light-electron microscopy to observe molecular dynamics in high resolution
Shouhei Kobayashi1, Masaaki Iwamoto1, Tokuko Haraguchi2
1Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan.
Microscopy (Oxford, England)
|July 8, 2016
Summary
Live correlative light-electron microscopy (CLEM) combines fluorescence and electron microscopy for high-resolution live-cell imaging. This technique enhances understanding of molecular functions within cellular structures for biology and medicine.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Molecular Imaging
Background:
- Fluorescence microscopy (FM) offers molecular specificity in live cells but lacks high spatial resolution.
- Electron microscopy (EM) provides ultrastructural detail but is limited to fixed, non-living samples.
- A gap exists in visualizing molecular dynamics at high resolution within living cells.
Purpose of the Study:
- To review methods combining live-cell fluorescence imaging with electron microscopy (live correlative light-EM or live CLEM).
- To highlight the advancements in high-resolution live-cell imaging.
- To underscore the utility of live CLEM in basic biology and medical research.
Main Methods:
- Live correlative light-EM (CLEM) integrates live-cell FM with subsequent in situ fixation and EM.
- Adaptations of FM techniques are applied within the CLEM workflow.
- Review of established and modified live CLEM protocols.
Main Results:
- Live CLEM enables simultaneous molecular selectivity and high-resolution structural imaging in living cells.
- Various fluorescence techniques can be integrated to enhance CLEM capabilities.
- The reviewed methods provide unprecedented insights into cellular structures and molecular functions.
Conclusions:
- Live CLEM significantly advances the understanding of cellular structure and molecular function.
- This technique bridges the resolution gap between FM and EM for live-cell studies.
- Live CLEM holds substantial promise for applications in both fundamental biological research and clinical medicine.
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