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Super-Resolution Live Cell Imaging of Subcellular Structures
Published on: January 13, 2021
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Spontaneously Blinking Fluorescent Protein for Simple Single Laser Super-Resolution Live Cell Imaging
Yoshiyuki Arai1,2, Hiroki Takauchi2, Yuhei Ogami2
1The Institute of Scientific and Industrial Research , Osaka University , 8-1 Mihogaoka, Ibaraki , Osaka 567-0047 , Japan.
ACS Chemical Biology
|July 3, 2018
Summary
Researchers developed a novel fluorescent protein, SPOON, enabling simple super-resolution microscopy. This protein exhibits spontaneous blinking, simplifying single molecule localization microscopy (SMLM) imaging with just one laser.
Area of Science:
- Biophysics
- Microscopy
- Molecular Biology
Background:
- Super-resolution imaging overcomes optical diffraction limits in microscopy.
- Single molecule localization microscopy (SMLM) requires photoswitchable probes and complex systems.
- Existing methods often necessitate multiple lasers or intricate setups for live-cell imaging.
Purpose of the Study:
- To develop a simplified SMLM platform for super-resolution imaging.
- To engineer a fluorescent protein with intrinsic photoswitching capabilities.
- To enable super-resolution imaging using a single excitation laser.
Main Methods:
- Development of a novel fluorescent protein, termed SPOON.
- Characterization of SPOON's photoswitching properties: light-induced off-switching and thermally induced on-switching.
- Implementation of SPOON in a single-molecule localization microscopy setup utilizing a single 488 nm laser.
Main Results:
- SPOON exhibits unique 'spontaneous' blinking behavior due to its switching mechanism.
- This blinking facilitates SMLM by enabling localization of individual molecules.
- A simplified SMLM platform was successfully demonstrated using only SPOON and a single laser.
Conclusions:
- The SPOON fluorescent protein offers a straightforward approach to SMLM.
- Its light- and thermally-controlled blinking simplifies super-resolution microscopy.
- This advancement lowers the technical barrier for advanced live-cell super-resolution imaging.
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