Two-photon MINFLUX with doubled localization precision

Zhao，Kun1,2; Xu，Xinzhu1,2; Ren，Wei1; Jin，Dayong3; Xi，Peng1,3,4

2022-12-01

10.1186/s43593-021-00011-x

eLight   影响因子和分区

2097-1710

2662-8643

Achieving localization with molecular precision has been of great interest for extending fluorescence microscopy to nanoscopy. MINFLUX pioneers this transition through point spread function (PSF) engineering, yet its performance is primarily limited by the signal-to-background ratio. Here we demonstrate theoretically that two-photon MINFLUX (2p-MINFLUX) could double its localization precision through PSF engineering by nonlinear effect. Cramér-Rao Bound (CRB) is studied as the maximum localization precision, and CRB of two-photon MINFLUX is halved compared to single-photon MINFLUX (1p-MINFLUX) in all three dimensions. Meanwhile, in order to achieve same localization precision with 1p-MINFLUX, 2p-MINFLUX requires only 1/4 of fluorescence photons. Exploiting simultaneous two-photon excitation of multiple fluorophore species, 2p-MINFLUX may have the potential for registration-free nanoscopy and multicolor tracking.

MINFLUX Single-molecule localization Super-resolution microscopy Two-photon microscopy

英语

通讯

National Natural Science Foundation of China[31971376];National Natural Science Foundation of China[61729501];National Natural Science Foundation of China[62025501];Natural Science Foundation of Beijing Municipality[JQ18019];Shenzhen Science and Technology Innovation Program[KQTD20170810110913065];

2-s2.0-85127325540

Scopus

1.Department of Biomedical Engineering,College of Future Technology,Peking University,Beijing,100871,China
2.Wallace H. Coulter Department of Biomedical Engineering,Georgia Institute of Technology and Emory University,Atlanta,30332,United States
3.UTS-SUStech Joint Research Centre for Biomedical Materials & Devices,Department of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Guangdong,Shenzhen,518055,China
4.National Biomedical Imaging Center,Peking University,Beijing,100871,China

Zhao，Kun,Xu，Xinzhu,Ren，Wei,et al. Two-photon MINFLUX with doubled localization precision[J]. eLight,2022,2(1).

Zhao，Kun,Xu，Xinzhu,Ren，Wei,Jin，Dayong,&Xi，Peng.(2022).Two-photon MINFLUX with doubled localization precision.eLight,2(1).

Zhao，Kun,et al."Two-photon MINFLUX with doubled localization precision".eLight 2.1(2022).