Fast and robust 3D MINFLUX excitation with a variable phase plate
bioRxiv (Cold Spring Harbor Laboratory)(2023)
摘要
MINFLUX has achieved record 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 around a fluorophore within a microsecond 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 with record speed, which we envision as an integral component of a high-performance and cost-effective open-source MINFLUX.
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关键词
robust 3d minflux excitation,phase
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