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题名	利用SOFI方法提高光片荧光显微镜横向分辨率的研究
作者	安坤
文献子类	硕士
导师	刘军
关键词	光片荧光显微镜 Light sheet fluorescence microscopy (LSFM) SOFI方法 Super-resolution optical fluctuation imaging algorithm (SOFI) 横向分辨率 Lateral resolution 斑马鱼 Zebrafish
其他题名	Improving lateral resolution of light sheet fluorescence microscopy with an enhanced SOFI
英文摘要	在过去的几十年里，随着生物科学研究的高速发展，越来越多的生物活体研究需要活体的实时高时空分辨率的成像技术。光片荧光显微成像是一种快速、低光漂白，和低光毒性的3D活体荧光成像方法。相比于其他的超分辨率荧光成像方法，该技术的光漂白和光毒性很低，并具有快速成像的优势，非常适用于有机活体的三维成像。然而，由于光片荧光显微镜需要正交放置的激发和成像物镜的特殊结构，导致其物镜的数值孔径相对于其他显微镜而言比较小，从而限制了光片荧光显微镜的横向分辨率。为了突破这个限制，进一步提高光片荧光显微成像的横向分辨率，本论文主要开展了以下几个方面的研究工作： 1. 搭建了一套双端泵浦的光片荧光显微镜装置，显微镜可在 488nm和405nm的激发光源之间切换，通过对斑马鱼和量子点等样品进行扫描采集成像，并对显微系统的性能进行了测试。 2. 我们将光片荧光显微镜技术和超分辨率光涨落成像（SOFI）算法相结合来提高光片荧光显微镜的横向分辨率。并且，我们对SOFI算法进行了改进，通过减少采集的图像数量，提高成像速度。在新算法中，加入的两个小波滤波器分别在时间和空间上消除低频背景噪声和原始图像读出噪声。利用商用的锑化镉量子点扩散到琼脂糖溶液和斑马鱼中作为样品，在我们自主搭建的光片荧光显微镜使用改进的SOFI算法，在不改变光片荧光显微镜的任何结构情况下，我们获得了约两倍横向分辨率的提高。; In the past decades, with the rapid development of biological research, more and more researches on living organisms require high spatial and temporal resolution imaging techniques. Light Sheet Fluorescence Microscopy (LSFM) is a fast 3D live fluorescence imaging method with low photo bleaching and photo toxicity. Compared with other super-resolution fluorescence imaging methods, this method has the advantages of low photo bleaching and photo toxicity as well as high imaging speed, which is very suitable for 3D imaging of living organisms. However, the lateral resolution of LSFM is usually limited by the NA of detection objective which could not be as high as other microscopies due to the two-orthogonal-objective setup of LSFM. In order to improve the situation, this thesis covers the following research works: 1. We built a double-side pumped LSFM which can be switched between the excitation light source 488nm and 405nm. Stage scanning mode for Zebrafish and quantum dots samples was achieved which prove the applicable of the setup. 2. We improve the lateral resolution of the fluorescence microscopy by combining the LSFM and the super-resolution light fluctuation imaging (SOFI). In addition, we improve the SOFI algorithm by reducing the number of images collected to improve the imaging speed. In the new algorithm, the two wavelet filters are used to eliminate the background noise and the noise of the original image in time and space. We use commercial cadmium antimonide quantum dots diffused into agarose solution and zebrafish as samples. The improved SOFI algorithm is used in our home-built light sheet fluorescence microscopy. We obtain a two fold increase in lateral resolution without any change in the structure of the fluorescence microscopy.
学科主题	光学工程
内容类型	学位论文
源URL	[http://ir.siom.ac.cn/handle/181231/30917]
专题	中国科学院上海光学精密机械研究所
作者单位	中国科学院上海光学精密机械研究所
推荐引用方式 GB/T 7714	安坤. 利用SOFI方法提高光片荧光显微镜横向分辨率的研究[D].

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