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BP: Fachverband Biologische Physik

BP 17: Bioimaging

BP 17.6: Vortrag

Mittwoch, 20. März 2024, 11:15–11:30, H 2032

eSRRF: Super-resolution radial fluctuation is going 3DR. F. Laine1, •H. S. Heil2, S. Coelho2, J. Nixon-Abell3, A. Jimenez4, T. Wiesner4, D. Martinez2, T. Galgani5, L. Régnier5, A. Stubb6, G. Follain6, S. Webster7, J. Goyette7, A. Dauphin5, A. Salles8, S. Culley1, G. Jacquemet6, B. Hajj5, C. Leterier4, and R. Henriques21UCL, London, UK — 2IGC, Oeiras, PT — 3Cambridge University, Cambridge, UK — 4CNRS-AMU, Marseille, FR — 5Institut Curie, Sorbonne Université, Paris, FR — 6University of Turku and Åbo Akademi University, Turku, FI — 7University of New South Wales, Sydney, AU — 8Institut Pasteur, Université de Paris, Paris, FR

Fluctuation-based image reconstruction extracts super-resolution details from brief wide-field image sequences, even under low light conditions (Gustafsson, Nat. Com. 2016). In the latest eSRRF version (Laine & Heil, Nat. Methods 2023, 10.1038/s41592-023-02057-w), reconstruction fidelity is significantly improved, with automated parameter optimization for image fidelity and resolution, crucial for avoiding artifacts (Culley, Nat. Methods 2018) and reducing user bias. While eSRRF excels in 2D super-resolution across microscopy techniques and biological systems, we've expanded its capability to 3D imaging.

For high-fidelity 3D live-cell nanoscopy with eSRRF, simultaneous detection of fluorescence fluctuations across multiple focal planes is crucial, achieved through a multifocus microscope (MFM, Hajj, PNAS 2014). This method allows volumetric super-resolution imaging of live cells at ~ 1 vol./s.

Keywords: Super-resolution Microscopy; Open source; Live-cell 3D imaging; Mulitfocus microscopy; Super-resolution Radial Fluctuation Microscopy

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