The automated measurement of foci in fluorecence microscopy

与丹麦技术大学的Flemming汉森博士的实验室合作，我们开发了一种全自动细胞识别和荧光焦点测量程序，该程序以图像分析程序Image Pro Plus中的宏的形式。这在检测大肠杆菌细胞中质粒和特定染色体位点形成的荧光灶的基本任务中非常成功。随着我们对荧光图像分析的需求越来越高，我们继续开发和调整该系统以满足这些需求。我们开发了一种分析工具“Inspect”，可以方便地分析具有双色荧光信息的图像。结合数据收集程序，我们开发了一系列电子表格程序来分析和描述数据。我们已经完成了图像分析软件开发的一个重要的新的推力。这允许自动识别处于各种分裂状态的多类细胞，以及正在分裂的病灶等。然后，这些细胞可以直接作为提取用于检查的图像的子类进行检查。然后可以定义和分析新的子类，并在数值数据和单个细胞图像之间进行无缝切换。这使我们能够比以前更详细地分析大量人口。今年，我们扩大了自动对焦测量和分析软件的实用性。该系统现在能够记录每个焦点的强度和纵横比。这允许评估DNA基因座的复制状态，并指示分离是否处于其最早阶段。此外，我们正在重新编写软件，以促进三重标记细胞的分析和双和三重标记的共定位的自动化检测。

In collaboration with the laboratory of Dr, Flemming Hansen, Technical University of Denmark, we have developed a fully automated cell recognition and fluorescent focus-measuring program in the form of a macro within the image analysis program Image Pro Plus. This has been very successful in the basic tasks of detecting fluorescent foci formed by plasmids and specific chromosomal loci in cells of the bacterium Escherichia coli. As our needs for analysis of fluorescent images become more demanding, we continue to develop and adapt this system to meet them. We have developed an analytical tool "Inspect" that facilitates the analysis of images that have two color fluorescence information. In conjunction with the data collection programs, we have developed an array of spreadsheet programs to analyze and portray the data. We have completed a major new thrust in the development of the image analysis software. This allows the automated identification of multiple classes of cells in various states of division, and of foci that are dividing etc. These cells can then be inspected directly as a sub-class of images extracted for inspection. New sub-classes can then be defined and analyzed with seamless switching between the numerical data and the individual cell images. This allows us to analyze large populations in far more detail than was previously possible. This year we have expanded the utility of our automated focus measuring and analysis software. The system is now capable of recording the intensity and aspect ratio of each focus. This allows assessment of the replicational state of the DNA locus and indicates whether segregation is in its earliest phase. In addition we are re-writing the software to facilitate the analysis of triple-labeled cells and the automated detection of co-localization of double and triple labels.