Enhancing the resolution of STED microscopy by SPLIT and flimGANE

Yuansheng Sun; Giorgio Tortarolo; Yuan-I Chen; Yin-Jui Chang; Ulas C. Coskun; Shih-Chu (Jeff) Liao; Beniamino Barbieri; Giuseppe Vicidomini; Hsin-Chih Yeh

doi:10.1117/12.3002745

12 March 2024 Enhancing the resolution of STED microscopy by SPLIT and flimGANE

Yuansheng Sun, Giorgio Tortarolo, Yuan-I Chen, Yin-Jui Chang, Ulas C. Coskun, Shih-Chu (Jeff) Liao, Beniamino Barbieri, Giuseppe Vicidomini, Hsin-Chih Yeh

Author Affiliations +

Proceedings Volume 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; 1284704 (2024) https://doi.org/10.1117/12.3002745
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Stimulated emission depletion (STED) microscopy is a powerful super-resolution microscopy technique that enables observation of sub-cellular structures with spatial resolution well below the diffraction limit. The higher the STED beam intensity, the higher the resolution, but at the cost of increased photo-damage, which significantly limits the application of STED microscopy in live specimens. The separation by lifetime tuning (SPLIT) technique uses a time-resolved acquisition and a phasor-based analysis to efficiently distinguish photons emitted from the center and from the periphery of the effective fluorescent region, thus improves the resolution of STED microscopy without increasing the STED beam intensity. Furthermore, the SPLIT method is combined with a deep learning-based phasor analysis algorithm termed flimGANE (fluorescence lifetime imaging based on a generative adversarial network), to improve the robustness of SPLIT-STED allowing improving the resolution up to 1.45 folds at the half of the depletion laser beam intensity.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Yuansheng Sun, Giorgio Tortarolo, Yuan-I Chen, Yin-Jui Chang, Ulas C. Coskun, Shih-Chu (Jeff) Liao, Beniamino Barbieri, Giuseppe Vicidomini, and Hsin-Chih Yeh "Enhancing the resolution of STED microscopy by SPLIT and flimGANE", Proc. SPIE 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, 1284704 (12 March 2024); https://doi.org/10.1117/12.3002745

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