三维显微图像复原及点扩散函数的研究

三维显微图像去卷积计算量巨大,运算时间长。评价和分析了三维点扩散函数、散焦像、光学切片以及不同三维点扩散函数去卷积的复原图像,进一步证明了散焦光信息对焦面像的干扰,主要来自于焦面像两侧附近的散焦像。提出了折中选择三维PSF的空间大小,可以获得良好的复原效果,并且减少运行时间。提出了频谱图均值的新概念,作为评价图像清晰及模糊程度的标准,并运用于散焦像、光学切片以及复原结果图的评价。     

Deconvolution of background-subtracted shift-and-add image by a modeled point-spread-function

Shift-and-add (SAA) is a simple method for celestial speckle imaging. However, the raw SAA-image, a direct output of the SAA operations, is not useful, because a seeing-dependent huge background is superimposed on the high-resolution image of the object. To obtain the latter, a background subtraction (BGS) is applied on the raw SAA-image. The BGS-image so obtained includes negativities due to an over-subtraction that causes brightnesses of the object’s image biased downward. The negativity in the BGS-image can be removed by a deconvolution with a point-spread-function (PSF) that has negative values. In this paper, we model the PSF with a shape that possesses peak and concave portions, and perform a deconvolution by iteratively estimating the object’s image and the model parameters. The simulated experiment has shown that the present algorithm can restore the object’s image with unbiased brightnesses. Processing the observed speckle data of Io (a Jupiter satellite) by the present method has yielded a feasible Io image with reduced negativities.     

Application of multi-frame approach in single-frame blind deconvolution

A new single-frame blind deconvolution algorithm for the linear shift-invariant imaging system is presented. The algorithm processes the partial images segmented from one single degraded image by multi-frame approach to recover the point spread function (PSF). Then a deconvolution method is employed to restore the whole image with the recovered PSF. In addition, in order to improve the fidelity and resolution of the recovered PSF, the coprimeness of the partial images is utilized. Results of simulated and real atmospheric turbulence degraded images using the algorithm are reported.     

谱域光学相干层析系统中基于解卷积方法的像质优化

谱域光学相干层析（spectral domain optical coherence tomography, 简记为SD-OCT）系统的轴向点扩散函数（point spread function, 简记为PSF）并不具备空不变特性,无法直接应用于解卷积运算.为实现SD-OCT系统成像质量基于解卷积算法的优化,本文采用数值校正后的轴向扫描信号和轴向有效PSF来实施基于Lucy-Richardson算法的解卷积运算,进而实现了SD-OCT系统中图像质量尤其是轴向分辨率的改善.本文理论分析了SD-OCT系统中导致轴向有效PSF随成像深度增大而下降和图像模糊的因素,阐述了利用解卷积算法实现图像质量优化的过程,基于建立的SD-OCT系统实施了不同成像深度位置处PSF的标定,并利用离散轴向位置处PSF的峰值拟合了轴向有效PSF的调制函数.利用调制函数对所有轴向扫描信号进行数值校正,然后根据轴向有效PSF进行解卷积算法.典型样品的解卷积图像重建结果表明,提出的解卷积方法能有效提高系统的轴向分辨率,同时有效抑制系统灵敏度随成像深度增大而下降的趋势. 关键词： 谱域光学相干层析 Lucy-Richardson解卷积 有效点扩散函数     

谱域光学相干层析系统中基于解卷积方法的像质优化

谱域光学相干层析（spectral domain optical coherence tomography, 简记为SD-OCT）系统的轴向点扩散函数（point spread function, 简记为PSF）并不具备空不变特性,无法直接应用于解卷积运算.为实现SD-OCT系统成像质量基于解卷积算法的优化,本文采用数值校正后的轴向扫描信号和轴向有效PSF来实施基于Lucy-Richardson算法的解卷积运算,进而实现了SD-OCT系统中图像质量尤其是轴向分辨率的改善.本文理论分析了SD-OCT系统中导致轴向有效PSF随成像深度增大而下降和图像模糊的因素,阐述了利用解卷积算法实现图像质量优化的过程,基于建立的SD-OCT系统实施了不同成像深度位置处PSF的标定,并利用离散轴向位置处PSF的峰值拟合了轴向有效PSF的调制函数.利用调制函数对所有轴向扫描信号进行数值校正,然后根据轴向有效PSF进行解卷积算法.典型样品的解卷积图像重建结果表明,提出的解卷积方法能有效提高系统的轴向分辨率,同时有效抑制系统灵敏度随成像深度增大而下降的趋势.     

Chromatin condensation and sensitivity of DNA in situ to denaturation during cell cycle and apoptosis--a confocal microscopy study

The goal of this study was to construct high resolution 3D confocal images of regions of condensed and extended chromatin in cell nuclei and individual chromosomes. It has been shown previously that sensitivity of DNA in situ to denaturation correlates with chromatin condensation and varies during cell cycle and apoptosis. Thus, detection of DNA which was partially denatured in situ provided a means to image areas of condensed chromatin. DNA denaturation was detected using a metachromatic dye acridine orange (AO) which differentially stains single stranded (ss) and double-stranded (ds) DNA sections. Early studies of denaturability of cellular DNA utilized flow cytometry and standard fluorescence microscopy. These techniques could not reveal small local differences in DNA denaturability within cell nucleus or in individual chromosomes. For instance, it was not possible to detect the initial points of chromosome condensation in G2-phase of the division cycle or in apoptosis. In order to achieve this goal we have recently extended these studies by applying confocal microscopy. We investigated DNA denaturability in normal human fibroblasts and HL-60 leukemic cells, at different stages of cell cycle and apoptosis. Following removal of RNA and partial denaturation of DNA with acid cells were stained with AO. Green (530 nm) and red (640 nm) fluorescence (exc. 457 nm) of non-denatured and denatured DNA was imaged by confocal microscopy. Blind deconvolution was used to further improve the quality of 3D images. Photobleaching of AO fluorescence was minimized and a correction for chromatic aberration and register shift was implemented. Nuclei of interphase cells exhibited predominantly green fluorescence representing AO binding to ds DNA. Punctuate areas of red fluorescence representing AO binding to denatured DNA and most likely associated with local regions of condensed chromatin were also present in all interphase nuclei. The proportion of denatured DNA increased in cells entering mitosis. In prophase individual condensing chromosomes exhibited varied proportions of green and red fluorescence indicating different content of denatured chromatin. In some chromosomes bands of denatured and denaturation-resistant chromatin were clearly resolved. In metaphase and anaphase chromosomes exhibited red fluorescence along all length of their arms indicating the highest and uniform susceptibility to denaturation. In telophase chromosomes contained predominantly denaturation-resistant DNA again and denaturated regions were significantly less abundant. At cytokinesis some decondensing chromosomes were still resolved. At this stage almost all regions of denatured DNA were located close to nuclear envelope. These regions may correspond to pockets of heterochromatin reforming at nuclear periphery. In early apoptosis condensation of chromatin appeared to commence in several distinct regions within nucleus. Some apoptotic bodies contained condensed chromatin surrounding central regions of extended chromatin. At late stages of apoptosis the whole volume of apoptotic bodies was occupied by condensed chromatin.     

自适应光学图像非对称图像迭代盲复原算法

 为了提高自适应光学图像复原效果,提出了一种新的多重约束非对称图像迭代盲解卷积算法。首先,在点扩散函数（PSF）频率域引入带宽有限约束来提高迭代盲解卷积算法的可靠性;然后,在PSF空间域引入支持域动态更新的思想以加快迭代盲解卷积算法收敛速度;最后,自动计算迭代盲解卷积算法的非对称因子以提高算法的自适应性。模拟实验结果表明,与RL-IBD算法比较,新算法迭代次数减少22.4%、峰值信噪比提高10.18 dB。在FK5-857和某双星的自适应光学图像复原实验中,也取得很好的复原效果。     

A two-stage method to correct aberrations induced by slide slant in bright-field microscopy

To achieve optimal image quality in bright field microscopy, the slide surface should be perpendicular to the optical axis of the microscope. However, in the recently proposed “slanted scan” slide acquisition technique, scan speed is increased by purposely slanting the slide by a small angle (of 3–5°) so that multiple focal depths can be imaged simultaneously. In this case, the slanted slide introduces a bend in the point spread function (PSF), resulting in a coma and other aberrations that degrade image quality. In this paper, we propose a two-stage deconvolution method specifically designed to correct the aberrations induced by a slanted scan, but with general applicability to high-resolution bright-field microscopy. Specifically, we initially apply phase deconvolution to correct the dominating coma aberration, before applying a conventional semi-blind deconvolution method to further improve image resolution and contrast. We also propose a novel method to estimate the degree of coma aberration and the PSF of the optics utilising actual cytology specimens. The efficacy of the proposed algorithm is demonstrated quantitatively on simulated data, against a ground-truth (object) image, and qualitatively on cervical cytology specimens. Results demonstrate both improved convergence speed of the two-stage approach, especially when correcting the bend in the PSF, and a resultant image quality that is comparable to a conventionally (flat) scanned specimen.     

4Pi confocal microscopy with alternate interference

We introduce 4Pi confocal microscopy with destructive interference of converging waves. Linear lobe deconvolution as well as nonlinear restoration of 4Pi confocal raw data with their point-spread functions (PSF's) leads to almost-identical images, irrespective of whether the 4Pi confocal PSF relies on constructive or destructive interference. Three-dimensional imaging of microtubules of a mouse fibroblast cell yielded an axial resolution near 100-nm in both cases. Moreover, restoration of 4Pi confocal images of the same object with alternate phases is introduced as a powerful test for (nonlinear) image restoration.     

Segmentation and counting of FISH signals in confocal microscopy images

In this paper we have presented a semi-automatic method for segmenting and counting the Fluorescence In Situ Hybridization (FISH) signals per cell nucleus in a 3D tissue image. The number of FISH signals indicate the gain (trisomy) or loss (monosomy) of certain base-sequences in deoxyribonucleic acid (DNA). The quantitative evaluation of the loss or gain in DNA is necessary in qualitative diagnostic molecular pathology. Multispectral volumetric images are obtained using the Confocal Microscope. Each consists of a red channel depicting the 3D morphology of the tissue and green channel containing the FISH signals. The red channel tissue image is segmented first to determine the membership of the FISH signal to a particular nuclei. Various segmentation methods starting from simple local thresholding to volume growing and active volumes are used for segmentation. A brief comparative study of the visual signal count by pathologist and our automatic count is also presented.     

Gradient-Based Blind Deconvolution with Phase Spectral Constraints

This paper proposes a new blind deconvolution method with additional phase spectral constraints for a blurred image. A degradation of an original image is mathematically modeled by a convolution of an original image and a point-spread function (PSF). The proposed method consists of the following three steps: (i) projection onto a complex set satisfying the phase spectral constraint in a frequency space; (ii) minimization of a cost function preserving the constrained phase spectra; and (iii) projection onto an image space satisfying nonnegative and support constraints. This method restores both the original image and the PSF with high accuracy. The effectiveness of the proposed method is verified by applying it to some blind deconvolution problems for digital images, and the experimental results show that the performance is superior to the conventional blind deconvolution methods.     

Blind image deconvolution using the zero-lag slice of higher-order statistics

A novel algorithm for blind image deconvolution using the zero-lag slice (ZLS) of higher-order statistics only is presented. This method first estimates the point-spread function (PSF) using the ZLS of its third-order moment (TOM) and then uses it with one of the known classical image deconvolution methods. The proposed method has simple computations for PSF estimation because it solves a nonlinear problem by using an iterative method with fast convergence. In each iteration, one need only calculate the ZLS of the TOM and estimate the PSF using simple two-dimensional operations. Furthermore, the method presented achieves good results, since the ZLS estimate obtained from the degraded image exhibits high reliability. The good performance of the proposed algorithm is demonstrated by applying it to synthetic and real data sets.     

用神经网络鉴别退化图像的模糊类型

提出了一种用神经网络鉴别退化图像的模糊类型的方法。由于采用不同降质方法得到退化图像的频谱差异较大,以此作为判别依据,用概率神经网络实现了对四种模糊类型:离焦,矩形,运动和高斯模糊的鉴别。根据神经网络的鉴别结果决定点扩散函数的初始估计值,可大大地提高盲解恢复算法的复原质量和系统点扩散函数的估计精度,扩大了算法的实用范围。     

Estimation of the ocular point spread function by retina modeling

Retinal imaging often suffers from blurring aberrations. With knowledge of the blurring point spread function (PSF), better images can be reconstructed by deconvolution techniques. We demonstrate a method to enhance the contrast of retinal cells by estimating the ocular PSF. This is done by finding the cells' positions and their intensity distributions and using these as a model for the image. The feasibility of this method is demonstrated by Wiener deconvolution both for adaptively and nonadaptively corrected images.     

采用色差先验约束的像差校正技术

本文通过分析自然图像的边缘3个通道之间的关联性,提出"同一物体的边缘在3个颜色通道应处于相同位置"的色差先验约束,该约束在数学上近似为各通道的相对导数相等,基于此色差先验约束,建立了一种新的像差校正模型即图像解卷积模型,并给出了基于交替方向乘子法的模型求解算法。实验结果表明:本文的像差校正技术可以提升图像的峰值信噪比10 dB以上,明显优于目前主流的BM3D和YUV算法,并且视觉提升效果明显,基本满足普通光学系统对像差的校正要求。     

基于目标和点扩展函数联合估计的点源目标图像近视解卷积

在传统的基于波前探测的解卷积方法中,由波前探测得到的点扩展函数被认为是精确的,并用维纳滤波进行复原,但是点扩展函数不可避免地存在误差,所以最终的复原目标图像质量不佳.为了解决该难题,提出了基于目标和点扩展函数联合估计的图像近视解卷积算法.它运用了点扩展函数和目标的先验信息,对点扩展函数和目标进行了规整和进一步约束,从而得到更优的恢复图像质量.对该方法的原理和实现过程进行了阐述,并将其运用于室内点源目标数据中.实验结果证明,与维纳滤波方法相比,该方法使图像恢复的效果得到明显改善.     

The use of a consumer grade photo camera in optical-digital correlator for pattern recognition and input scene restoration

In this work an optical-digital correlator for pattern recognition and input scene restoration is described. Main features of the described correlator are portability and ability of multi-element input scenes processing. The correlator consists of a consumer grade digital photo camera with a diffractive optical element (DOE) inserted as a correlation filter. Correlation of an input scene with a reference image recorded on the DOE are provided optically and registered by the digital photo camera for further processing. Using obtained correlation signals and DOE’s point spread function (PSF), one can restore the image of the input scene from the image of correlation signals by digital deconvolution algorithms.The construction of the correlator based on the consumer grade digital photo camera is presented. The software procedure that is necessary for images linearization of correlation signals is described. Experimental results on optical correlation are compared with numerical simulation. The results of images restoration from conventionally and specially processed correlation signals are reported. Quantitative estimations of accuracy of correlation signals as well as restored images of the input scene are presented.     

空间变化PSF非盲去卷积图像复原法综述

传统的图像复原一般认为点扩散函数(PSF)是空间不变的,实际光学系统由于受到像差等因素的影响,并非严格的线性空间不变系统,基于空间变化PSF的非盲去卷积图像复原法逐渐体现其优越性。空间变化PSF的非盲去卷积图像复原法先准确估计图像空间变化的PSF,再利用非盲去卷积算法对图像进行复原,有利于恢复出高质量图像。本文从算法的角度综述了近几年提出的基于空间变化PSF的非盲去卷积图像复原方法,并对比了基于强边缘预测估计PSF的非盲去卷积法、基于模糊噪声图像对PSF估计非盲去卷积法等算法的优缺点,各算法分别在PSF估计精确度、振铃效应抑制效果、适用范围等方面体现出各自的优劣。空间变化PSF的非盲去卷积图像复原法的研究,有利于推进图像复原技术向更高水平发展,使光学系统往轻小型化方向发展,从而在多个科学领域发挥其重要作用。     

NaYF4∶Eu3+, Tm3+, Yb3+材料中Stokes和反Stokes发光研究

合成了Eu3+,Tm3+和Yb3+掺杂的NaYF₄材料。360 nm光激发呈蓝色发光,峰值位于452 nm,对应Tm3+的1D₂→3F₄跃迁;395 nm光激发呈橙色发光,峰值位于591 nm,对应Eu3+的5D₀→7F₁跃迁;409 nm光激发呈红色发光,峰值位于613 nm,对应Eu3+的5D₀→7F₂跃迁;980 nm光激发呈蓝色和红色发光,发光峰位于474和646 nm。蓝光来源Tm3+的1G₄ →3H₆跃迁,红光来源Tm3+的1G₄→3F₄跃迁。在双对数曲线中,蓝光474 nm和红光646 nm的斜率分别为2.1和2.4,在980 nm光激发下,蓝光和红光发射都是双光子过程。还研究了材料的吸收光谱,并利用X射线衍射,扫描电镜测试了材料的物相结构和微观形貌。结果表明：NaYF₄∶Eu3+, Tm3+, Yb3+材料具有较规则的六方相结构,结晶良好。     

利用维纳滤波改善声透镜光声成像系统的分辨率

为了克服衍射效应对光声成像系统分辨率的限制,需要采用逆卷积方法进行图像反演.从理论上分析了声透镜成像原理,模拟仿真了声透镜的点扩展函数对声透镜成像系统分辨率的影响和维纳滤波解卷积方法复原光声成像的过程,并利用自搭建的声透镜光声成像系统进行了深入的实验研究,得到了物平面上相距4 mm和3 mm的两个黑胶带点的直接成像光声...