RESTORATION OF CONFOCAL IMAGES FOR QUANTITATIVE IMAGE-ANALYSIS
RESTORATION OF CONFOCAL IMAGES FOR QUANTITATIVE IMAGE-ANALYSIS
复制标题
DOI：
10.1111/j.1365-2818.1995.tb03593.x
复制
发表时间：
1995-05-01
期刊：
影响因子：
--
通讯作者：
STRASTERS, KC
中科院分区：
文献类型：
--
作者：
VANDERVOORT, HTM;STRASTERS, KC
通过引入显微体积成像技术，最值得注意的是共聚焦荧光显微镜（CFM），微观物体的三维（3-D）结构的定量研究已经成为可能。虽然CFM是一个真正的体积成像仪，其特定的成像特性引起的图像失真，并妨碍随后的定量分析。因此，在分析之前恢复共焦图像是先决条件。失真可以分为几类：由于自吸收，漂白效应，几何效应和由于衍射效应的失真在图像中的区域的衰减。其中，吸收和衍射效应是最重要的。本文介绍了一种校正衍射畸变的方法。讨论了恢复共焦图像所需的所有步骤，包括一种常规测量仪器性能的新方法。为了测试恢复过程，恢复荧光平面对象的图像。结果表明，在z分辨率和没有振铃伪影相当大的改善。图像分析的方法的相关性证明了应用3-D纹理分析的合成对象的恢复和未恢复的图像的结果的比较。此外，该方法可以成功地应用于生物对象，如间期细胞核的噪声荧光图像。
Quantitative studies of three-dimensional (3-D) structure of microscopic objects have been made possible through the introduction of microscopic volume imaging techniques, most notably the confocal fluorescence microscope (CFM). Although the CFM is a true volume imager, its specific imaging properties give rise to distortions in the images and hamper subsequent quantitative analysis. Therefore, it is a prerequisite that confocal images are restored prior to analysis. The distortions can be divided into several categories: attenuation of areas in the image due to self-absorption, bleaching effects, geometrical effects and distortions due to diffraction effects. Of these, absorption and diffraction effects are the most important. This paper describes a method aimed at the correction of diffraction-induced distortions. All the steps necessary in restoring confocal images are discussed, including a novel method to measure instrumental properties on a routine basis. To test the restoration procedure an image of a fluorescent planar object was restored. The results show a considerable improvement in the z-resolution and no ringing artefacts. The relevance of the method for image analysis is demonstrated by a comparison of results of applying 3-D texture analysis to restored and unrestored images of a synthetic object. Furthermore, the method can be successfully applied to noisy fluorescence images of biological objects, such as interphase cell nucei.