受激辐射损耗超分辨成像技术研究

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。

Super resolution imaging technology of stimulated emission depletion

Stimulated emission depletion (STED) is a kind of super resolution fluorescence microscopy imaging technology. It can break through the traditional optical diffraction limit, and make the far field optical resolution improved to less than 100 nm, which is widely used in biomedical field, and becomes one of the hotspots in optical imaging research. A STED microscope system based on ultra-continuous spectrum picoseconds pulsed white laser source was introduced, and super-resolution imaging was realized. The optimized results were discussed from the aspects of precise collimation, pulse delay and residual intensity of the STED light, therefore the best imaging effect was obtained. The experiments data of the about 25 nm diameter fluorescent microspheres imaging shows that the resolution of the system is about 60 nm, which is much higher than the diffraction limit. In addition, the system succeeds in realizing super resolution imaging of nuclear pore complexes, microtubules and microfilaments and a series of biological samples. Some obscure structures imaged in confocal microscopy can be legible in STED imaging.