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Accurate Background Subtraction in STED Nanoscopy by Polarization Switching
- Accurate Background Subtraction in STED Nanoscopy by Polarization Switching
- Lee, Jong-Chan; Ma, Ye; Han, Kyu Young; Ha, Taekjip
- DGIST Authors
- Lee, Jong-Chan
- Issue Date
- ACS Photonics, 6(7), 1789-1797
- Article Type
- Author Keywords
- super-resolution microscopy; STED nanoscopy; background noise; background suppression; optical resolution
- STED nanoscopy helped usher in resolution revolution of optical microscopy and has been widely used to study subcellular structures and dynamics. To achieve optical resolution beyond the diffraction limit, STED nanoscopy employs 10(4)-10(5) times more intense light compared to conventional imaging methods, giving rise to additional background that can be detrimental to imaging and quantification of dense samples. Here, we report a simple and easy-to-implement method to suppress background noise by polarization switching (psSTED). STED imaging requires a well-defined polarization state of a STED laser (typically left-circular polarization). By switching to right-circular polarization, the spatial profile of the STED laser changes to fill up the center of the donut-shaped beam, enabling an accurate background recording. Through a straightforward subtraction of the recorded background from the regular STED image, we achieved background-subtracted STED imaging. With simulation and experiment, we demonstrate that psSTED outperforms other background subtraction methods.
- American Chemical Society
- Related Researcher
Single-molecule Biophysics and Advanced Bioimaging Laboratory
Liquid-liquid phase separation; Super-resolution imaging;
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- Department of New BiologySingle-molecule Biophysics and Advanced Bioimaging Laboratory1. Journal Articles
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