光学成像系统中有限孔径对光波的衍射,使得光学显微成像技术的分辨率受到"衍射极限"限制而无法进一步提高.自1873年E.K.Abbe提出该问题以来,衍射极限就一直是学术界研究的热点.近年来,随着高强度激光、高灵敏探测器等光电器件研制技术以及新型荧光探针开发等相关领域的快速发展,光学显微技术衍射极限问题的研究迎来了新的契机,超分辨显微成像技术(super-resolution microscopy,SRM)在近十年内取得了令人瞩目的巨大成就.本文从空域和频域角度回顾了衍射极限分辨率的基本原理,并据此对目前常见的各种SRM技术"绕过"衍射极限提高分辨率的机理给予了详解,同时介绍了各类技术的发展动态和研究方向;作为SRM的一个新的重要的发展趋势,本文详细介绍了超分辨关联显微技术的最新研究进展,包括SRM与活细胞实时荧光显微、荧光寿命显微、光谱测量和成像、电子显微、原子力显微、质谱技术等的关联,着重讨论了各类超分辨关联显微技术的作用和意义;最后,对SRM技术和超分辨关联显微技术的未来发展方向进行了展望.%The diffraction of the finite aperture in the optical imaging system restricts further improvement of the resolution of optical microscopy, which is called the "diffraction limit". Since raised by Ernst Abbe in 1873, the problem of diffraction limit has been one of the foci of academic research. In recent years, with the rapid development of related fields such as the development of optoelectronic devices including high energy lasers and high sensitivity detectors and the development of new fluorescent probes, the problem of diffraction limit in optical microscopy ushered in a new opportunity, and super-resolution microscopy (SRM) has made remarkable achievements in the past decade. The basic principles of diffraction limited resolution in both space and frequency domains are reviewed, and on this basis, the mechanisms for the various SRM technologies to circumvent the diffraction limit and improve the resolution are explained in detail. The development trends and research directions of various SRM techniques are also introduced. As a new and important development trend of SRM, correlative super-resolution microscopy and its recent progress are reviewed, including correlative studies on SRM and time-lapse live cell fluorescence microscopy, fluorescence lifetime imaging microscopy, spectrometry and spectroscopy, electron microscopy, atomic force microscopy, etc. The role and significance of various correlative super-resolution microscopy are discussed. The future development of super-resolution microscopy and correlative super-resolution microscopy is also prospected.
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