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介绍了四能级系统的受激辐射耗尽(STED)荧光显微镜的基本原理。按照时间顺序介绍了面包圈型耗尽光焦斑的实现方法。对于横向分辨率的改善,最初使用了光路偏移法,后来发展到均分相位板法,再到成熟的螺旋相位板法;另外还介绍了半波相位板法对轴向分辨率的改善。从宽带光源激发、连续光源激发、多色多通道、快速成像和双光子激发等方面综述了受激辐射耗尽荧光显微镜实验装置的逐步完善。最后展望了受激辐射耗尽荧光显微镜的发展前景。
The basic principle of stimulated emission depletion (STED) fluorescence microscopy for four-level systems is introduced. According to the chronological order, the method of realizing the torch-type depleted light focal spot is introduced. For the improvement of lateral resolution, the optical path shift method was first used, and then the phase-division method and then to the mature spiral phase method. The improvement of the axial resolution by the half-wave phase plate method was also introduced. The experimental equipment of stimulated emission depleted fluorescence microscopy was reviewed in the aspects of broadband light source excitation, continuous light source excitation, multi-color multi-channel, rapid imaging and two-photon excitation. Finally, the prospect of stimulated depletion fluorescence microscopy was prospected.