Phase aberration compensation of digital holographic microscopy based on least squares surface fitting

Digital holographic microscopy allows the numerical reconstruction of the complex wavefront of samples, especially biological samples such as living cells. In digital holographic microscopy, a microscope objective is introduced to improve the transverse resolution of the sample; however a phase aberration in the object wavefront is also brought along, which will affect the phase distribution of the reconstructed image. We propose here a numerical method to compensate for the phase aberration of thin transparent objects with a single hologram. The least squares surface fitting with points number less than the matrix of the original hologram is performed on the unwrapped phase distribution to remove the unwanted wavefront curvature. The proposed method is demonstrated with the samples of the cicada wings and epidermal cells of garlic, and the experimental results are consistent with that of the double exposure method.     

Complex-amplitude-based phase unwrapping method for digital holographic microscopy

A complex-amplitude-based phase unwrapping approach for digital holographic microscopy is proposed in this paper. A quality map is derived directly from the reconstructed complex amplitude distribution of object wave to evaluate noise influence and phase reliability in the wrapped phase image. Quality-guided phase unwrapping algorithm is then implemented with the quality map to retrieve continuous phase profile. Unwrapping errors caused by unreliable phase data are successfully suppressed. The effectiveness of this method is demonstrated with simulation and experimental results.     

Physical spherical phase compensation in reflection digital holographic microscopy

Reflection configured digital holographic microscopy (DHM) can perform accurate optical topography measurements of reflecting objects, such as MEMs, MOEMs, and semiconductor wafer. It can provide non-destructive quantitative measurements of surface roughness and geometric pattern characterization with nanometric axial resolution in real-time. However, the measurement results may be affected by an additional phase curvature introduced by the microscope objective (MO) used in DHM. It needs to be removed either by numerical compensation or by physical compensation.We present a method of physical spherical phase compensation for reflection DHM in the Michelson configuration. In the object arm, collimated light is used for illumination. Due to the use of the MO, the object wavefront may have a spherical phase curvature. In the reference arm, a lens and mirror combination is used to generate a spherical recording reference wave in order to physically compensate the spherical phase curvature of the object wavefront. By controlling the position of the mirror and the sample stage, the compensation process has been demonstrated. The relative positions of the test specimen and the reference mirror must be fixed for the physical spherical phase to be totally compensated. A numerical plane reference wave is preferred for the numerical reconstruction of the phase introduced by the test specimen. Experimental results on wafer pattern recognition are also given.     

Phase aberration compensation by spectrum centering in digital holographic microscopy

Firstly, an approach is proposed for eliminating the tilt phase aberration introduced by the tilt reference wave. It is based on the argument distribution of the hologram spectrum to locate exactly the position of the carrier frequency of the virtual image. This tilt aberration will be corrected by shifting the filtered hologram spectrum to the coordinate origin of the frequency domain, without knowledge of the focal length of the imaging lens or distances in the setup. Then the subsequent quadratic phase aberration compensation is performed only by adjusting a single parameter. The method is demonstrated by the phase contrast imaging of the cervical carcinoma cells.     

基于Zernike多项式进行波面拟合研究

基于Zernike多项式利用Householder变换法对不同空间频率径向误差和不同口径局部误差进行了拟合,得到了拟合误差的均方根(RMS)值,分析了Zernike多项式的局限性。结果表明,Zernike在拟合径向误差时,受到最大空间频率的限制;在拟合局部误差时要受到局部误差口径大小的限制,并且会引起额外的波动。     

Phase reconstruction in lensless digital in-line holographic microscopy

The phase reconstruction in a digital in-line holographic microscopy is compared using two numerical reconstruction methods. The first method uses one Fourier transform and second one uses three Fourier transforms. It is shown that the latter method gives improved object phase reconstruction as compared to the former.     

泽尼克多项式校正全息阵列光镊像差的实验研究

像差会影响光镊对粒子的捕获效果. 全息阵列光镊中, 像差不仅来自光学元件, 由特定算法设计的光阱相位片也会在光路中引入像差. 本文通过液晶空间光调制器加载泽尼克多项式相位图, 对全息阵列光镊中由光栅透镜组型算法引起的像差进行校正. 结果显示: 利用三阶泽尼克多项式可有效消除光路中由光栅透镜组型算法引 起的慧差, 使得捕获2 μm聚苯乙烯小球的阵列光阱刚度提高了约40%; 对比不同项的像差校正结果发现, 全息阵列光镊中由算法引起的慧差 与光学元件引起的像差一样, 也会对阵列光阱的捕获效果产生较大影响; 同时根据一阶像差校正结果可得光栅透镜 组型算法对于一阶泽尼克像差具有鲁棒性. 实验结果表明, 对全息阵列光镊中由 算法引起的像差进行校正, 对于提高光阱的捕获效果和深化对算法特性的认识都具有重要意义.     

Zernike多项式拟合曲面中拟合精度与采样点数目研究

为了研究采样点数目对由Zernike多项式所拟合的曲面的拟合精度的影响,采用不完全归纳法,取得了采样点与拟合精度之间的数据关系。结果表明:不同的测试函数遵循相同的规律,即采样点数目达到一定数目后,拟合精度随采样点的变化很小。并且,通过计算得到了在较高拟合精度时,采样点数目与Zernike多项式的项数之间的变化规律,实际例子证明了该变化规律的正确性,其对于Zernike多项式拟合曲面具有很好的指导意义。     

Disturbance-free digital holographic microscopy via a micro-phase-step approach

This work proposes a cost-effective, simple, micro-phase-step (MPS) method for suppressing the zero-order diffraction and conjugate-image interferences that are caused during digital holographic microscopic image reconstruction. The proposed MPS method replaces the conventional phase modulation approach; it uses a rotatable cover glass that enables smooth modification of the incidence angle and the optical path of the reference beam. This setup allows the phase step to be accurately estimated by shifting the reference wave phase more freely close to π/2, at which the background noise can be suppressed more effectively. In the proposed MPS method, the optimal conditions for suppressing conjugate-image interference are identified using a relatively moderate intensity distribution and suppression of noise in the numerically reconstructed object wave-field. In addition, the proposed method mitigates the effect of disturbances that are caused by environmental factors, such as minor vibrations and small changes in temperature and humidity. Importantly, only two holograms are required to satisfy the objective of image reconstruction. The results in this work reveal that even with intentional interference caused by minor vibrations, conjugate-image interference can still be suppressed by determining the phase deviation between the two original holograms.     

环形大口径平面镜圆Zernike多项式拟合精度分析

采用瑞奇-康芒实验装置和利用圆Zernike多项式分别对不同遮拦比环域进行了拟合。结果表明,在遮拦比小于0.4的环域上,用圆Zernike多项式拟合,残差PV和残差RMS值不是很大;在遮拦比大于0.4的环域上,需用环Zernike多项式拟合才能保证一定的拟合精度。     

Rough surface characterization using off-axis digital holographic microscopy compensated with self-hologram rotation

In this paper, an off-axis digital holographic microscopy compensated with self-hologram rotation is presented. The process is implemented via subtracting the unwrapped phase maps of the off-axis parabolic hologram and its rotation 180° to eliminate the tilt induced by the angle between the spherical object wave O and the plane reference wave R. Merit of the proposed method is that it can be done without prior knowledge of physical parameters and hence can reconstruct a parabolic hologram of 1024 × 768 pixels within tens of milliseconds since it doesn't require a digital reference wave. The method is applied to characterize rough gold bumps and the obtained results were compared with those extracted from the conventional reconstruction method. The comparison showed that the proposed method can characterize rough surfaces with excellent contrast and in real-time. Merit of the proposed method is that it can be used for monitoring smaller biological cells and micro-fluidic devices.     

Parallel-quadrature phase-shifting digital holographic microscopy using polarization beam splitter

We present a digital holography microscopy technique based on a parallel-quadrature phase-shifting method. Two π/2 phase-shifted holograms are recorded simultaneously using polarization phase-shifting principle, slightly off-axis recording geometry, and two identical CCD sensors. The parallel phase-shifting is realized by combining circularly polarized object beam with a 45° degree polarized reference beam through a polarizing beam splitter. DC term is eliminated by subtracting the two holograms from each other and the object information is reconstructed after selecting the frequency spectrum of the real image. Both amplitude and phase object reconstruction results are presented. Simultaneous recording eliminates phase errors caused by mechanical vibrations and air turbulences. The slightly off-axis recording geometry with phase-shifting allows a much larger dimension of the spatial filter for reconstruction of the object information. This leads to better reconstruction capability than traditional off-axis holography.     

Study on the simplified phase-shifting digital holographic microscopy

In this paper, a new simplified technique for effectively eliminating the zero order and the conjugate virtual image in digital holographic microcopy, which makes use of two-step phase-shifting method of just recording two holograms and an intensity image of object wave, is proposed. Meanwhile, combined with the principle of making full use of spatial bandwidth of the CCD sensor by in-line lens-less Fourier holographic recording geometry, the theory and experimental methods to increase the resolution of the reconstructed image in digital holography by using phase-shifting technique are detailedly analyzed. At end, the validity and availability of this technique has been demonstrated through the off-axis and in-line Fourier transform recording geometry. The study provides some theoretical and experimental guidance for the design and operation of a digital holographic microscopy system.     

Common-path phase-shifting digital holographic microscopy: A way to quantitative phase imaging and superresolution

We present an experimental setup useful for complex amplitude evaluation and phase image quantification of three-dimensional (3-D) samples in digital holographic microscopy (DHM). It is based on a common-path interferometric configuration performed by dividing the input plane in two contiguous regions and by placing a translation grating near to the Fourier plane. Then, complex amplitude distribution of the sample under test is recovered with phase-shifting standard method obtained by moving the grating using a linear motion stage. Some experimental results of an USAF resolution test are presented for different numerical aperture (NA) microscope lenses. In a second part, the proposed setup is tested under superresolution purposes. Based on the object’s spectrum shift produced by off-axis illumination, we use time multiplexing to generate a synthetic aperture enlargement that improves the final image resolution. Experimental results for the case of a biosample (human red blood cells) and a commercial low NA microscope lens validates the suggested superresolution approach.     

High resolution digital holographic microscopy for the study of aggregated natural cellulose nanowhisker fibers

In this paper, digital holographic (DH) microscopy demonstrates its ability to perform a full characterization of nanofibers. The high resolution and magnification of the presented method to study the nanofibers are tested using standard MIL-STD-150A 1951 USAF resolution test target. In this investigation, aggregated natural cellulose nanowhisker fibers are positioned in the front of the microscopic objective using a 3D translation stage in the object arm of DH setup. The recorded off-axis holograms are refocused using the angular spectrum method. The reconstructed complex field is used to calculate optical phase and intensity distributions of the object at different reconstruction depths. A simple algorithm is used to define the focused image with suitable accuracy. The dimensions and orientation of the fibers can be evaluated from the optical field at different depths. Then, the shape and textures along the aggregated natural cellulose nanowhisker fiber can be presented in a 3D space.     

An experimental investigation of three-dimensional particle aggregation using digital holographic microscopy

The tendency of particles to aggregate depends on particle-particle and particle-fluid interactions. These interactions can be characterized but it requires accurate 3D measurements of particle distributions. We introduce the application of an off-axis digital holographic microscopy for measuring distributions of dense micrometer (2 μm) particles in a liquid solution. We demonstrate that digital holographic microscopy is capable of recording the instantaneous 3D position of particles in a flow volume. A new reconstruction method that aids identification of particle images was used in this work. About 62% of the expected number of particles within the interrogated flow volume was detected. Based on the 3D position of individual particles, the tendency of particle to aggregate is investigated. Results show that relatively few particles (around 5–10 of a cohort of 1500) were aggregates. This number did not change significantly with time.     

Reconstruction algorithms applied to in-line Gabor digital holographic microscopy

This paper investigates the application of Fresnel based numerical algorithms for the reconstruction of Gabor in-line holograms. We focus on the two most widely used Fresnel approximation algorithms, the direct method and the angular spectrum method. Both algorithms involve calculating a Fresnel integral, but they accomplish it in fundamentally different ways. The algorithms perform differently for different physical parameters such as distance, CCD pixel size, and so on. We investigate the constraints for the algorithms when applied to in-line Gabor digital holographic microscopy. We show why the algorithms fail in some instances and how to alter them in order to obtain useful images of the microscopic specimen. We verify the altered algorithms using an optically captured digital hologram.     

Error compensation for mirror symmetry absolute test

A method is proposed to compensate intrinsic error in mirror symmetry absolute test. Because of the limitation of rotation times of flat A, intrinsic error of CNθ terms occurs in reconstructed wavefronts of flats A, B and C. If flat A is rotated to a new azimuthal position, and the wavefront difference between two measurements before and after rotation is calculated, the Zernike coefficients of CNθ terms can be obtained by solving coefficient equations due to rotation invariability of the form of Zernike polynomials in polar coordinates. Therefore, the intrinsic error of CNθ terms may be compensated. Because the amount of CNθ terms is infinite, the compensated terms are decided in terms of the balance between intrinsic error reduction and computational effort. Computer simulation proves the validity of the proposed method.     

基于Zernike多项式表示色域边界的色域映射

在彩色图像的跨媒体复制中,由于不同媒体有不同的色域,所以在大多数情况下色域映射是不可避免的。不论采用何种映射算法,色域映射的第一步是确定有关媒体的色域边界。目前大多数的色域边界都是用一组测量的或用其它方法得到的离散数据来表示的。然而,由于这种表示方式是利用一组离散数据来实施色域映射的两个基本处理(即确定任一指定的等色调面平面中色域的边缘线以及确定色域边界与一条映射线的交点),比较复杂和费时,因此希望找到一种色域边界的解析表达式来解决这一问题。简述了不久前提出的一种解析函数色域边界表示方法,并用实例说明了如何利用这种解析表示(Zernike多项式)来实现上述的边缘线确定和交点确定。最后给出了几种映射算法的色域映射完整过程及其结果。     

Performance analysis of quantitative phase retrieval method in Zernike phase contrast X-ray microscopy

Since the invention of Zernike phase contrast method in 1930,it has been widely used in optical microscopy and more recently in X-ray microscopy.Considering the image contrast is a mixture of absorption and phase information,we recently have proposed and demonstrated a method for quantitative phase retrieval in Zernike phase contrast X-ray microscopy.In this contribution,we analyze the performance of this method at different photon energies.Intensity images of PMMA samples are simulated at 2.5 keV and 6.2 keV,respectively,and phase retrieval is performed using the proposed method.The results demonstrate that the proposed phase retrieval method is applicable over a wide energy range.For weakly absorbing features,the optimal photon energy is 2.5 keV,from the point of view of image contrast and accuracy of phase retrieval.On the other hand,in the case of strong absorption objects,a higher photon energy is preferred to reduce the error of phase retrieval.These results can be used as guidelines to perform quantitative phase retrieval in Zernike phase contrast X-ray microscopy with the proposed method.