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Super-Resolution Live Cell Imaging of Subcellular Structures
Published on: January 13, 2021
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Multicolor re-scan super-resolution imaging of live cells
Jiuling Liao1, Longchao Chen1, Xianyuan Xia1
1Research Laboratory for Biomedical Optics and Molecular Imaging, Shenzhen Key Laboratory for Molecular Imaging, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
Quantitative Imaging in Medicine and Surgery
|July 9, 2019
Summary
This study introduces a new multicolor re-scan confocal microscopy (RCM) system for super-resolution imaging. The advanced RCM overcomes spatial mismatch issues, enabling clearer visualization of cellular structures in live cells.
Area of Science:
- Cell biology
- Microscopy
- Biophysics
Background:
- Conventional multicolor fluorescence microscopy is limited by diffraction in imaging subcellular organelles.
- Re-scan confocal microscopy (RCM) offers super-resolution but previous methods suffered from spatial mismatch.
- Sequential laser scanning in multicolor RCM causes image misalignment.
Purpose of the Study:
- To develop a novel RCM system for multicolor super-resolution imaging.
- To overcome spatial mismatch issues in multicolor RCM.
- To enable high-resolution imaging of live cells with multiple fluorophores.
Main Methods:
- Implemented a spectrograph as the multicolor detection system.
- Utilized a linear spectral unmixing algorithm for fluorophore separation.
- Applied gamma correction to restore image resolution after reconstruction.
Main Results:
- Achieved a lateral resolution of approximately 171 nm, a 1.8-fold improvement over wide-field imaging.
- Successfully resolved spectrally overlapping fluorophores using three types of fluorescent beads.
- Demonstrated suitability for imaging multicolor-labeled organelles in live cells.
Conclusions:
- The novel RCM system provides multicolor super-resolution images of live cells without spatial mismatch, photobleaching, or photodamage.
- This system offers a new tool for observing dynamic molecular and organelle interactions in cells.
- Potential for advancing studies on cellular dynamics and molecular interactions.
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