三维显微图像的分辨率评估方法及应用  

Resolution Evaluation Method and Applications of 3D Microscopic Images

作　　者：罗喜亮 周舟 黄江枫 董香江 郑刚 付玲 Luo Xiliang;Zhou Zhou;Huang Jiangfeng;Dong Xiangjiang';Zheng Gang;Fu Ling(Brittoyi Chance Center for Biomedical Photonics,Wuhan National Laboratory for Optoelectronics,Huazhong University of Scieyice and Technology,Wuhan,Hubei 430074,China;MoE Key Laboratory for Biomedical Photonics,School of Engineering Sciences,Huazhong University of Science and Technology,Wuhan,430074,China;School of Electronic Information and Communications,Huazhong University of Science and Technology,Wuhan,Hubei 430074,China)

机构地区：[1]华中科技大学武汉光电国家研究中心BrittonChance生物医学光子学中心,湖北武汉430074 [2]华中科技大学工程科学学院生物医学光子学教育部重点实验室,湖北武汉430074 [3]华中科技大学电子信息与通信学院,湖北武汉430074

出　　处：《中国激光》2022年第5期139-149,共11页Chinese Journal of Lasers

基　　金：国家自然科学基金(61890952)。

摘　　要：现有分辨率评估方法,如瑞利判据、阿贝判据、半峰全宽方法等都具有一定的应用局限性。本文采用基于切片傅里叶壳层相关(sFSC)的频域分辨率评估方法来评估系统的实际成像能力,该方法对分辨率的评估结果仅取决于图像质量,不受系统成像理论的影响,是一种客观、直接的计算方法。它将傅里叶壳层分成楔形壳层对,每个选择器都是一对镜像楔形,以改善由三维荧光成像各向异性带来的分辨率评估问题。实验结果表明,sFSC可作为一种无参考三维图像分辨率评估方法,且利用sFSC方法的分辨率结果所拟合的三维高斯点扩展函数(PSF)进行图像反卷积操作,能有效恢复图像纹理细节,提高图像信噪比,且相比于其他PSF估计方法,sFSC方法具有更好的性能。Objective Selecting an appropriate method to analyze images obtained with similar imaging systems and with similar image resolution values but clearly different image quality is challenging.Traditional resolution evaluation methods,such as Rayleigh criterion,Abbe criterion,and FWHM,are not sufficiently reproducible to evaluate the resolution of an actual image and are not universal among different systems.Theoretically,the point spread function(PSF)size of laser scanning confocal and two-photon microscopes is fixed;however,the actual imaging resolution is affected by many other factors.SRM systems,such as SIM,STED,PALM,and STORM,achieve super-resolution by PSF modulation technology or single molecule positioning technology.However,the theoretical resolution of these systems is only related to the limited experimental conditions that can be achieved in practice,such as photobleaching,phototoxicity,and molecular positioning accuracy.Thus,there is no fixed resolution.Therefore,traditional resolution evaluation methods cannot be applied to all systems simultaneously.Here,we adopt a resolution evaluation method based on sectioned Fourier shell correlation(sFSC),which is an image-based method that does not require any a priori information and can be directly used to calculate the resolution of a given image.We expect that this method will be used to evaluate and compare the imaging capabilities of different imaging systems and can be applied to optimize image restoration.Methods We adopt a resolution evaluation method based on frequency domain correlation.First of all,we use the checkerboard method to divide the original image by pixel into two groups that have the same details but uncorrelated noise.Then,we apply a Fourier transform to obtain the spectrum diagram of all the images.After that,the corresponding strength values in the wedge shell selector are substituted to a correlation calculation formula until all frequency shells and azimuths are completed.In the next step,the cutoff frequency of each azimuth angle i

关 键 词：生物光学 无参考三维图像分辨率评估 切片傅里叶壳层相关 三维显微图像复原 反卷积 

分 类 号：Q334[生物学—遗传学]