Simple and robust 3D MINFLUX excitation with a variable phase plate

作     者：Takahiro Deguchi Jonas Ries Takahiro Deguchi;Jonas Ries

作者机构：European Molecular Biology LaboratoryCell Biology and BiophysicsHeidelbergGermany. Max Perutz LabsVienna Biocenter Campus(VBC)ViennaAustria. University of ViennaCenter for Molecular BiologyDepartment of Structural and Computational BiologyViennaAustria. University of ViennaFaculty of PhysicsViennaAustria 

出 版 物：《Light(Science & Applications)》 (光（科学与应用)（英文版）)

年 卷 期：2024年第13卷第7期

页      面：1354-1363页

核心收录：

学科分类：0401[教育学-教育学] 04[教育学] 0702[理学-物理学] 

基　　金：H2020 Marie Skłodowska-Curie Actions(RobMin grant no.101031734 to T.D.) the European Research Council(grant no.ERC CoG-724489 to J.R.) the European Molecular Biology Laboratory(T.D.and J.R.) 

主　　题：excitation phase switched 

摘      要：MINFLUX has achieved extraordinary resolution in superresolution imaging and single fluorophore *** is based on localizing single fluorophores by rapid probing with a patterned beam that features a local intensity *** implementations,however,are complex and expensive and are limited in speed and ***,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 *** 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 *** 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.