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Super-Resolution Imaging by Dual Iterative Structured Illumination Microscopy

Super-Resolution Imaging by Dual Iterative Structured Illumination Microscopy

来源:bioRxiv_logobioRxiv
英文摘要

Three-dimensional (3D) multicolor super-resolution imaging in the 50-100 nm range in fixed and living cells remains challenging. We extend the resolution of structured illumination microscopy (SIM) by an improved nonlinear iterative reconstruction algorithm that enables 3D multicolor imaging with improved spatiotemporal resolution at low illumination intensities. We demonstrate the performance of dual iterative SIM (diSIM) imaging cellular structures in fixed cells including synaptonemal complexes, clathrin coated pits and the actin cytoskeleton with lateral resolutions of 60-100 nm with standard fluorophores. Furthermore, we visualize dendritic spines in 70 μm thick brain slices with an axial resolution < 200 nm. Finally, we image dynamics of the endoplasmatic reticulum and microtubules in living cells with up to 255 frames/s.

Spindler Marie-Christine、Benavente Ricardo、Netz Ralf、Kleppe Dr. Ingo、L?schberger Anna、Klein Teresa、Sauer Markus、Novikau Yauheni

Department of Cell and Developmental Biology, Biocenter, University of W¨1rzburg, Am HublandDepartment of Cell and Developmental Biology, Biocenter, University of W¨1rzburg, Am HublandZEISS Research Microscopy Solutions, Carl Zeiss Microscopy GmbH, Advanced DevelopmentZEISS Research Microscopy Solutions, Carl Zeiss Microscopy GmbH, Advanced DevelopmentZEISS Research Microscopy Solutions, Carl Zeiss Microscopy GmbH, Advanced DevelopmentDepartment of Biotechnology and Biophysics, Biocenter, University of W¨1rzburg, Am HublandDepartment of Biotechnology and Biophysics, Biocenter, University of W¨1rzburg, Am HublandZEISS Research Microscopy Solutions, Carl Zeiss Microscopy GmbH, Advanced Development

10.1101/2021.05.12.443720

生物科学研究方法、生物科学研究技术生物物理学细胞生物学

super-resolution microscopystructured illumination microscopy3D multicolor super-resolution imaging

Spindler Marie-Christine,Benavente Ricardo,Netz Ralf,Kleppe Dr. Ingo,L?schberger Anna,Klein Teresa,Sauer Markus,Novikau Yauheni.Super-Resolution Imaging by Dual Iterative Structured Illumination Microscopy[EB/OL].(2025-03-28)[2025-04-28].https://www.biorxiv.org/content/10.1101/2021.05.12.443720.点此复制

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