受激辐射耗尽荧光显微镜的原理与实验装置研究进展

Principle and Progress of Experimental Setup on Stimulated Emission Depletion Fluorescence Microscope

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。

This paper describes an introduction in both setup and rate equations of stimulated emission depletion （STED） fluorescence microscope. The methods of generating STED beam and a series of phase plates used in STED fluorescence microscope are introduced. The development of the setup on STED fluorescence microscope, including broadband STED, continuous-wave （CW） STED, multi-color STED, fast STED and single-wavelength two-photon emission （SW2PE） STED, is described. Future prospects for the development of the STED microscope experimental setup is put forward; new laser sources, new phase plates, three-dimensional （3D） super-resolution, and a variety of imaging modalities combined imaging are the main development directions in the future.