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题名	菲涅尔非相干相关数字全息在光片显微成像中的研究与应用
作者	梁东
文献子类	硕士
导师	刘军
关键词	LSFM LSFM FINCH FINCH 偏振平面透镜 Polarization Directed Flat Lens 分辨率 Resolution
其他题名	Investigation and Application of Fresnel Incoherent Correlation Digital Holography in light sheet fluorescence microscopy
英文摘要	在过去的几十年里，伴随着生命科学研究的高速发展，共聚焦激光扫描显微镜对生命科学研究领域做出了巨大的贡献，然而其成像中的逐点扫描过程非常耗时，使得活体样本中的大多数生理学过程难以捕捉，并且有较大的光毒性。为了研究活体生物体内的微观动态过程，非接触性和低侵入性的显微镜技术则是一种必不可少的研究手段。光片荧光显微成像（Light Sheet Fluorescence Microscopy，LSFM）是一种快速、低光漂白和低光毒性的三维活体荧光成像方法。它通过选择激发生物样本的不同区域和控制激发层厚度来抑制背景荧光噪声，提高图像的对比度，并与高速探测器相结合实现快速宽场成像，具有低光漂白和低光毒性等众多优点，非常适合活体样本的三维成像。 数字全息显微术将显微成像技术和数字全息术相结合，实现了小体积生物样品的三维显微成像和物场振幅及相位的全场、非破坏性及高精度的定量测量与分析。传统的数字全息技术对光源的相干性要求较高，而非相干全息的出现降低了对该方面的要求，从而可以应用于非相干的荧光成像。在非相干全息中，菲涅尔非相干相关数字全息术(Fresnel Incoherent Correlation Digital Holography, FINCH)是一种非扫描、低噪声、高分辨的三维成像技术，在当前已成为国内外非相干数字全息的研究热点。FINCH技术基于物信息和菲涅尔波带片之间的相关实现非相干全息图的记录，利用相移法在重建过程中抑制孪生像的影响。它可以在非相干全息的基础上与任何标准的光学成像技术相匹配实现荧光显微成像。 在本论文中，我们将光片显微成像技术与菲涅尔非相干相关数字全息相结合，主要开展了以下几个方面的研究工作： 1、我们选择了一种新型的对偏振敏感的几何相位透镜(偏振定向平面透镜）代替传统FINCH技术中的空间光调制器作为全息记录光路中的分光器件来实现波前调制和衍射分光，自主设计并完成了完整的基于偏振定向平面透镜的菲涅尔非相干相关数字全息产生装置的搭建，验证了偏振定向平面透镜可代替空间光调制器实现非相干全息图的记录，这不仅克服了对空间光调制器的高性能和高衍射效率的要求，而且大大降低了实验装置的复杂性和高昂的成本，直线型结构使得该装置结构紧凑、搭建方便和易于调节。 2、我们将光片荧光显微技术与基于偏振定向平面透镜的菲涅尔非相干相关数字全息装置相结合，搭建了一套双端泵浦的LSFM-FINCH荧光显微镜装置，利用该装置中的LSFM实现了生物样本的荧光成像。; In the past decades, with the rapid development of life science researches, Confocal Laser Scanning Microscope has made tremendous contributions to the field of life science research, but the point-by-point scanning process in confocal imaging which is time-consuming and phototoxicity, most physiological processes in living samples are difficult to capture. In order to study the microscopic dynamic processes in living organisms, non-contact and minimally invasive microscope technology is an essential research method. Light Sheet Fluorescence Microscopy (LSFM) is a fast 3D live fluorescence imaging method with low photo bleaching and photo toxicity. It can select different areas of the biological sample to stimulate and control the thickness of the excitation layer to suppress the background fluorescence noise, improve the contrast of the image and combine with high-speed detectors to achieve fast wide-field imaging. It has many advantages such as low photobleaching and low phototoxicity, which is very suitable for three-dimensional imaging of living samples. Digital holographic microscopy combines microscopic imaging technology with digital holography to achieve three-dimensional microscopic imaging of a small-volume biological sample and full-field, non-destructive and high-precision quantitative measurement and analysis of the amplitude and phase of the object field. However, the traditional digital holography technology requires higher coherence of the light source, the appearance of incoherent holography reduces the requirements on this aspect, so that it can be applied to incoherent fluorescence imaging. In in-coherent holography, Fresnel Incoherent Correlation Digital Holography (FINCH) is a non-scanning, low-noise, high-resolution 3D imaging technology, which has become a research hotspot in incoherent holography research field. FINCH realizes the recording of in-coherent holograms based on the correlation between the object information and the fresnel zone plate. The phase-shifting method is used to suppress the influence of twin images during the reconstruction process. It can be matched with any standard optical imaging technology based on incoherent holography to achieve fluorescence microscopy imaging. In this paper, we combined light sheet microscopy with Fresnel Incoherent Correlation Digital Holography, and mainly carried out the following research work: 1. We had chosen a new geometric phase lens (polarization directed flat lens) which is sensitive to polarization, instead of the spatial light modulator in the traditional FINCH technology as a splitting device in the holographic recording optical path to realize wavefront modulation and diffraction splitting. The complete Fresnel Incoherent Correlation Digital Holography generation device based on polarization directed flat lens is completed after our independent design. It was verified that the polarization directed flat lens can replace the spatial light modulator to achieve recording of inherent holograms. It was not only overcome the high-performance requirement and high diffraction efficiency of the spatial light modulator, but also greatly reduced the complexity and high cost of the experimental device. Because of the linear structure, the device was compact and easy to adjust. 2. We combined the light sheet fluorescence microscopy with a Fresnel incoherent correlation digital holography device based on a polarization directed flat lens to set up a double-side pumped LSFM-FINCH fluorescence microscope device, and realized fluorescence imaging of biological samples.
学科主题	光学工程
内容类型	学位论文
源URL	[http://ir.siom.ac.cn/handle/181231/31037]
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	梁东. 菲涅尔非相干相关数字全息在光片显微成像中的研究与应用[D].

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