Smart 3D-FISH:: Automation of distance analysis in nuclei of interphase cells by image processing
Smart 3D-FISH:: Automation of distance analysis in nuclei of interphase cells by image processing
复制标题
DOI：
10.1002/cyto.a.20170
复制
发表时间：
2005-09-01
期刊：
影响因子：
3.7
通讯作者：
Boudier, T
中科院分区：
文献类型：
--
作者：
Gué, M;Messaoudi, C;Boudier, T
背景资料：在具有保留的三维核结构的细胞中通过荧光原位杂交（3D-FISH）检测荧光探针对于研究染色质的组织和间期核中基因的定位是有用的。快速和可靠的测量特定的亚染色体区域和基因的荧光斑点的相对定位，将提高对细胞结构和function.Methods的理解：3D-FISH协议，共聚焦显微镜，和数字图像analysis.Results：新的软件（智能3D-FISH）已开发出自动化的过程中的斑点分割和距离测量的图像从3D-FISH实验。它可以处理任何数量的荧光点并结合4 '，6-二氨基-2-苯基吲哚复染细胞核的图像，以测量细胞核中点位点的相对定位和点间距离。从一个试点实验中使用智能3D-FISH ENL，MLL，和AF 4基因在两个淋巴母细胞株的结果是令人满意的和一致的数据发表在literation.Conclusion：智能3D-FISH应大大方便图像处理和分析的3D-FISH图像提供了一个有用的工具，以克服繁重的任务，图像分割的基础上，用户自定义的参数和减少数据分析的主观性。它可以作为ImageJ软件的一组插件。(c)2005 Wiley-Liss，Inc.
Background: Detection of fluorescent probes by fluorescence in situ hybridization in cells with preserved three-dimensional nuclear structures (3D-FISH) is useful for studying the organization of chromatin and localization of genes in interphase nuclei. Fast and reliable measurements of the relative positioning of fluorescent spots specific to subchromosomal regions and genes would improve understanding of cell structure and function.Methods: 3D-FISH protocol, confocal microscopy, and digital image analysis were used.Results: New software (Smart 3D-FISH) has been developed to automate the process of spot segmentation and distance measurements in images from 3D-FISH experiments. It can handle any number of fluorescent spots and incorporate images of 4 ',6-diamino-2-phenylindole counterstained nuclei to measure the relative positioning of spot loci in the nucleus and inter-spot distance. Results from a pilot experiment using Smart 3D-FISH on ENL, MLL, and AF4 genes in two lymphoblastic cell lines were satisfactory and consistent with data published in the literature.Conclusion: Smart 3D-FISH should greatly facilitate image processing and analysis of 3D-FISH images by providing a useful tool to overcome the laborious task of image segmentation based on user-defined parameters and decrease subjectivity in data analysis. It is available as a set of plugins for ImageJ software. (c) 2005 Wiley-Liss, Inc.