Advanced Photonics Nexus, 2023, 2 (5): 056008, Published Online: Sep. 18, 2023
High-fidelity SIM reconstruction-based super-resolution quantitative FRET imaging
super-resolution structured illumination microscopy Förster resonance energy transfer living cells quantitative measurement
Abstract
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.
Zewei Luo, Guodong Zang, Ge Wu, Mengting Kong, Zhengfei Zhuang, Tongsheng Chen. High-fidelity SIM reconstruction-based super-resolution quantitative FRET imaging[J]. Advanced Photonics Nexus, 2023, 2(5): 056008.