首页|Simple and robust 3D MINFLUX excitation with a variable phase plate
Simple and robust 3D MINFLUX excitation with a variable phase plate
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MINFLUX has achieved extraordinary resolution in superresolution imaging and single fluorophore tracking.It is based on localizing single fluorophores by rapid probing with a patterned beam that features a local intensity minimum.Current implementations,however,are complex and expensive and are limited in speed and robustness.Here,we show that a combination of an electro-optical modulator with a segmented birefringent element such as a spatial light modulator produces a variable phase plate for which the phase can be scanned on the MHz timescale.Bisected or top-hat phase patterns generate high-contrast compact excitation point-spread functions for MINFLUX localization in the x,y,and z-direction,respectively,which can be scanned across a fluorophore within a microsecond,switched within 60 microseconds and alternated among different excitation wavelengths.We discuss how to compensate for non-optimal performance of the components and present a robust 3D and multi-color MINFLUX excitation module,which we envision as an integral component of a high-performance and cost-effective open-source MINFLUX.
Takahiro Deguchi、Jonas Ries
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European Molecular Biology Laboratory,Cell Biology and Biophysics,Heidelberg,Germany
Max Perutz Labs,Vienna Biocenter Campus(VBC),Vienna,Austria
University of Vienna,Center for Molecular Biology,Department of Structural and Computational Biology,Vienna,Austria
University of Vienna,Faculty of Physics,Vienna,Austria
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H2020 Marie Sk?odowska-Curie ActionsEuropean Research CouncilEuropean Molecular Biology Laboratory
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