Zewei Luo 1,2†Guodong Zang 1,2Ge Wu 1,2Mengting Kong 1,2[ ... ]Tongsheng Chen 1,2,*
Author Affiliations
1 South China Normal University, College of Biophotonics, MOE Key Laboratory of Laser Life Science, Guangzhou, China
2 South China Normal University, College of Biophotonics, Guangdong Key Laboratory of Laser Life Science, Guangzhou, China
Structured illumination-based super-resolution Förster resonance energy transfer microscopy (SIM-FRET) provides an approach to resolving molecular behavior localized in intricate biological structures in living cells. However, SIM reconstruction artifacts will decrease the quantitative analysis fidelity of SIM-FRET signals. To address these issues, we have developed a method called HiFi spectrum optimization SIM-FRET (HiFi-SO-SIM-FRET), which uses optimized Wiener parameters in the two-step spectrum optimization to suppress sidelobe artifacts and achieve super-resolution quantitative SIM-FRET. We validated our method by demonstrating its ability to reduce reconstruction artifacts while maintaining the accuracy of FRET signals in both simulated FRET models and live-cell FRET-standard construct samples. In summary, HiFi-SO-SIM-FRET provides a promising solution for achieving high spatial resolution and reducing SIM reconstruction artifacts in quantitative FRET imaging.
super-resolution structured illumination microscopy Förster resonance energy transfer living cells quantitative measurement 
Advanced Photonics Nexus
2023, 2(5): 056008

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