数字显微全息关键技术研究
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摘要
本文主要研究实现微小或显微相位物体的三维轮廓再现的数字显微全息关键技术。
论文基于数字显微全息技术,构建了多种数字离轴显微全息再现系统和光栅相移数字同轴显微全息再现系统。并实现了基于全息再现原理的相位物体,如相位光栅、活体细胞的三维相位再现。为获得正确的原始显微深度结构或轮廓再现结果,本文对数字显微全息技术展开深入研究。在此基础上,提出了以下四项创新点。
1、最佳再现距离d和数字再现光波倾斜因子k_x,k_y的调整技术
再现距离d和数字再现光波倾斜因子k_x,k_y是实现数字全息图正确数值再现的重要参数。离焦误差和倾斜误差的存在对再现信息尤其是再现相位信息均会引起畸变的结果。本文提出基于修正平均梯度的调焦方法,抑制了菲涅耳近似再现算法中再现像大小随再现距离变化对聚焦判断的影响,获得最佳再现距离。同时根据傅里叶变换的频移特性,实现了倾斜因子k_x,k_y的自动调整。
2、基于再现平面相位相减的误差校正技术
全息图数值再现信息中,系统误差的抑制或消除方法很多,但各有利弊。本文所提出的再现平面相位相减全息图处理新方法,利用样本全息图和背景全息图,通过再现相位相减处理,减去除样本之外的系统相位误差,实现再现平面上原始物波准确及高精度的相位再现结果。该方法能有效、合理地消除光源噪声、光学器件所引入的相位误差和其它背景相位误差,且保证了记录样本的相位信息中与误差分布类似的分项不被误处理。在活体细胞的全息再现中,该方法的优点体现尤为显著。
3、预放大数字离轴全息图中二次相位误差校正技术
显微物镜所引入的二次项相位误差的校正是完成预放大数字显微全息图数值再现的重要环节。针对现有校正方法的不足,本文所提出的基于显微物镜后焦面的二次项相位误差校正、衍射计算获得原始物波的预放大数字显微全息图数值再现的新方法,避免了硬件校正的严格限制,校正调整参数具有单一性,且只需单幅全息图再现,对记录样本特性也没有限制。
4、光栅相移数字同轴全息图处理技术
传统相移技术用于同轴全息再现,需要引入参考光源,从而无法实现单光束数字同轴全息的记录。基于点衍射理论的数字同轴全息再现技术仅限于高度透明和尺寸远小于记录介质面径尺寸的记录样本。本文提出了利用数字微镜器件DMD实现的光栅相移数字同轴全息再现技术。该技术利用正弦振幅光栅相移技术和衍射处理技术实现同轴全息图的数值再现,获得原始物波信息。该方法进一步结合DMD的分光特性,可实现光栅相移数字同轴全息正交再现系统,为实现显微物体的高精度形貌或结构再现指引了很好的方向和提供了有力保证。
In this dissertation, digital micro-holography for the reconstruction of three dimensional profiles of the small or micro phase objects and its key techniques are studied.
Based on digital micro-holography, several kinds of digital off-line micro-holographic systems are built and one kind of grating phase-shifting digital in-line micro-holographic system is presented to realize the reconstruction of three dimensional phase information of the tested phase objects including phase grating and living cell. In order to get the right reconstruction information of the original micro-structure or the profile, we have paid attention to the research of digital micro-holography, and propose four novel techniques as follows:
1.Estimation technique of optimal reconstruction distance d and titling factors k_x, k_y of the numerical reconstruction wave
The reconstruction distanced and the titling factors k_x, k_y of the numerical reconstruction wave are three important parameters to realize the right numerical reconstruction of digital hologram. Their aberrations will effect the right information reconstruction, specially the phase information reconstruction. The presented new estimation method of the optimal reconstruction distance based on the revised average grad method suppresses the effect of the lateral resolution changing with the reconstruction distance on the estimation of reconstruction distance to get the exact reconstruction distance. While we carry out the titling factor k_x, k_y computed automatically according to the frequency-shifting characteristic of Fourier transform.
2.Aberration correction technique based on phase subtraction method at the reconstruction plane
All kinds of the existing methods for the aberration correction of the reconstructed numerically phase of digital hologram have the different merits and disadvantages. We put forward a new phase subtraction method at the reconstruction plane, in which two holograms are digitally recorded and numerically reconstructed, respectively, and their reconstruction phase distributions are subtracted to reduce the phase aberration unwanted to get the exact original phase information. It can reduces the phase aberration from the light source, optical components and background, and while ensure that the phase of the recorded objects won't be subtracted mistakenly if some of the distributions of its sub-terms are similar with the phase aberration. The proposed method shows especially its advantages in the profile reconstruction of living cells.
3.Aberration correction technique for the quadratic phase of pre-magnified digital off-line micro-hologram
For pre-magnified digital micro-holography, the compensation of the quadratic phase produced by micro-objective is vital. Aiming to the disadvantages of the existing compensating methods we propose a new reconstruction method, in which firstly the quadratic phase is compensated at the back focal plane of micro-objective, and then get the original object wave-field by diffraction calculation. The proposed new method reveals some advantages, such as the additional optical parts does not be added, the parameter in numerical compensation process shows unique, and only single hologram is needed.
4.Processing technique of grating phase-shifting digital in-line hologram
In traditional phase-shifting in-line holography, the recording of hologram needs two beams because the reference light source must be added. The reconstruction of digital in-line hologram by single beam based on point diffraction theory has the limitations that the tested objects must be high transparent and their sizes need be less than one of the recording medium. We provide the grating phase-shifting digital in-line holography with Digital Micro-mirror Device(DMD). By taking advantage of the reflection and diffraction characteristics of DMD, it integrates the sine amplitude grating phase-shifting technique and diffraction processing technique into digital holography to realize the numerical reconstruction of in-line hologram, and then to get the original wave-field. Further, combining with the beam splitting characteristic of DMD, the grating phase-shifting digital in-line holographic orthogonal reconstruction system can be built, which provide the better way and feasibility for the high precision profile or structure reconstruction.
论文基于数字显微全息技术,构建了多种数字离轴显微全息再现系统和光栅相移数字同轴显微全息再现系统。并实现了基于全息再现原理的相位物体,如相位光栅、活体细胞的三维相位再现。为获得正确的原始显微深度结构或轮廓再现结果,本文对数字显微全息技术展开深入研究。在此基础上,提出了以下四项创新点。
1、最佳再现距离d和数字再现光波倾斜因子k_x,k_y的调整技术
再现距离d和数字再现光波倾斜因子k_x,k_y是实现数字全息图正确数值再现的重要参数。离焦误差和倾斜误差的存在对再现信息尤其是再现相位信息均会引起畸变的结果。本文提出基于修正平均梯度的调焦方法,抑制了菲涅耳近似再现算法中再现像大小随再现距离变化对聚焦判断的影响,获得最佳再现距离。同时根据傅里叶变换的频移特性,实现了倾斜因子k_x,k_y的自动调整。
2、基于再现平面相位相减的误差校正技术
全息图数值再现信息中,系统误差的抑制或消除方法很多,但各有利弊。本文所提出的再现平面相位相减全息图处理新方法,利用样本全息图和背景全息图,通过再现相位相减处理,减去除样本之外的系统相位误差,实现再现平面上原始物波准确及高精度的相位再现结果。该方法能有效、合理地消除光源噪声、光学器件所引入的相位误差和其它背景相位误差,且保证了记录样本的相位信息中与误差分布类似的分项不被误处理。在活体细胞的全息再现中,该方法的优点体现尤为显著。
3、预放大数字离轴全息图中二次相位误差校正技术
显微物镜所引入的二次项相位误差的校正是完成预放大数字显微全息图数值再现的重要环节。针对现有校正方法的不足,本文所提出的基于显微物镜后焦面的二次项相位误差校正、衍射计算获得原始物波的预放大数字显微全息图数值再现的新方法,避免了硬件校正的严格限制,校正调整参数具有单一性,且只需单幅全息图再现,对记录样本特性也没有限制。
4、光栅相移数字同轴全息图处理技术
传统相移技术用于同轴全息再现,需要引入参考光源,从而无法实现单光束数字同轴全息的记录。基于点衍射理论的数字同轴全息再现技术仅限于高度透明和尺寸远小于记录介质面径尺寸的记录样本。本文提出了利用数字微镜器件DMD实现的光栅相移数字同轴全息再现技术。该技术利用正弦振幅光栅相移技术和衍射处理技术实现同轴全息图的数值再现,获得原始物波信息。该方法进一步结合DMD的分光特性,可实现光栅相移数字同轴全息正交再现系统,为实现显微物体的高精度形貌或结构再现指引了很好的方向和提供了有力保证。
In this dissertation, digital micro-holography for the reconstruction of three dimensional profiles of the small or micro phase objects and its key techniques are studied.
Based on digital micro-holography, several kinds of digital off-line micro-holographic systems are built and one kind of grating phase-shifting digital in-line micro-holographic system is presented to realize the reconstruction of three dimensional phase information of the tested phase objects including phase grating and living cell. In order to get the right reconstruction information of the original micro-structure or the profile, we have paid attention to the research of digital micro-holography, and propose four novel techniques as follows:
1.Estimation technique of optimal reconstruction distance d and titling factors k_x, k_y of the numerical reconstruction wave
The reconstruction distanced and the titling factors k_x, k_y of the numerical reconstruction wave are three important parameters to realize the right numerical reconstruction of digital hologram. Their aberrations will effect the right information reconstruction, specially the phase information reconstruction. The presented new estimation method of the optimal reconstruction distance based on the revised average grad method suppresses the effect of the lateral resolution changing with the reconstruction distance on the estimation of reconstruction distance to get the exact reconstruction distance. While we carry out the titling factor k_x, k_y computed automatically according to the frequency-shifting characteristic of Fourier transform.
2.Aberration correction technique based on phase subtraction method at the reconstruction plane
All kinds of the existing methods for the aberration correction of the reconstructed numerically phase of digital hologram have the different merits and disadvantages. We put forward a new phase subtraction method at the reconstruction plane, in which two holograms are digitally recorded and numerically reconstructed, respectively, and their reconstruction phase distributions are subtracted to reduce the phase aberration unwanted to get the exact original phase information. It can reduces the phase aberration from the light source, optical components and background, and while ensure that the phase of the recorded objects won't be subtracted mistakenly if some of the distributions of its sub-terms are similar with the phase aberration. The proposed method shows especially its advantages in the profile reconstruction of living cells.
3.Aberration correction technique for the quadratic phase of pre-magnified digital off-line micro-hologram
For pre-magnified digital micro-holography, the compensation of the quadratic phase produced by micro-objective is vital. Aiming to the disadvantages of the existing compensating methods we propose a new reconstruction method, in which firstly the quadratic phase is compensated at the back focal plane of micro-objective, and then get the original object wave-field by diffraction calculation. The proposed new method reveals some advantages, such as the additional optical parts does not be added, the parameter in numerical compensation process shows unique, and only single hologram is needed.
4.Processing technique of grating phase-shifting digital in-line hologram
In traditional phase-shifting in-line holography, the recording of hologram needs two beams because the reference light source must be added. The reconstruction of digital in-line hologram by single beam based on point diffraction theory has the limitations that the tested objects must be high transparent and their sizes need be less than one of the recording medium. We provide the grating phase-shifting digital in-line holography with Digital Micro-mirror Device(DMD). By taking advantage of the reflection and diffraction characteristics of DMD, it integrates the sine amplitude grating phase-shifting technique and diffraction processing technique into digital holography to realize the numerical reconstruction of in-line hologram, and then to get the original wave-field. Further, combining with the beam splitting characteristic of DMD, the grating phase-shifting digital in-line holographic orthogonal reconstruction system can be built, which provide the better way and feasibility for the high precision profile or structure reconstruction.
引文
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