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受激辐射损耗显微成像(STED)是一种超分辨荧光显微成像技术,它能够突破传统光学衍射极限的限制,把远场光学分辨率提高到百纳米以内,被广泛应用于生物医学等领域,是目前光学显微成像领域研究的热点之一。采用了一种基于超连续谱皮秒脉冲白激光光源的STED显微系统,实现超分辨成像。并从精密合束、脉冲延迟和损耗光残留光强几个方面探讨系统优化,从而获得最佳的成像效果。对直径约25 nm荧光微球成像实验的数据表明:该系统成像分辨率可达约60 nm,分辨能力远远高于衍射极限。另外,系统成功实现了对核孔复合物、微管和微丝等一系列生物样品的超分辨成像,共聚焦成像中某些模糊不清的结构在STED成像中清晰可辨。
Stimulated radiation depletion microscopy (STED) is a super-resolution fluorescence microscopy technique that overcomes the limitations of traditional optical diffraction limits and increases far-field optical resolution to within 100 nanometers. It is widely used in biomedical and other fields Field, is one of the hot spots in the field of optical microscopy. A STED microscopy system based on supercontinuum picosecond pulse white laser light source was used to realize super-resolution imaging. The system optimization is discussed in terms of precision beam combining, pulse delay and residual light intensity loss, so as to obtain the best imaging results. The experimental data of the fluorescent microspheres with a diameter of about 25 nm show that the imaging resolution of the system is about 60 nm and the resolving power is much higher than the diffraction limit. In addition, the system successfully achieves super-resolution imaging of a series of biological samples such as nuclear pore complexes, microtubules and microfilaments. Some obscure structures in the confocal imaging are clearly identifiable in STED imaging.