Oeck Sebastian, Malewicz Nathalie M, Hurst Sebastian, Al-Refae Klaudia, Krysztofiak Adam, Jendrossek Verena
Institute of Cell Biology (Cancer Research), University of Duisburg-Essen, Medical School, Virchowstrasse 173, 45122 Essen, Germany.
Radiat Res. 2017 Jul;188(1):114-120. doi: 10.1667/RR14746.1. Epub 2017 May 11.
The quantitative analysis of foci plays an important role in various cell biological methods. In the fields of radiation biology and experimental oncology, the effect of ionizing radiation, chemotherapy or molecularly targeted drugs on DNA damage induction and repair is frequently performed by the analysis of protein clusters or phosphorylated proteins recruited to so called repair foci at DNA damage sites, involving for example γ-H2A.X, 53BP1 or RAD51. We recently developed "The Focinator" as a reliable and fast tool for automated quantitative and qualitative analysis of nuclei and DNA damage foci. The refined software is now even more user-friendly due to a graphical interface and further features. Thus, we included an R-script-based mode for automated image opening, file naming, progress monitoring and an error report. Consequently, the evaluation no longer required the attendance of the operator after initial parameter definition. Moreover, the Focinator v2-0 is now able to perform multi-channel analysis of four channels and evaluation of protein-protein colocalization by comparison of up to three foci channels. This enables for example the quantification of foci in cells of a specific cell cycle phase.
病灶的定量分析在各种细胞生物学方法中发挥着重要作用。在放射生物学和实验肿瘤学领域，电离辐射、化疗或分子靶向药物对DNA损伤诱导和修复的影响，通常通过分析招募到DNA损伤位点所谓修复病灶的蛋白质簇或磷酸化蛋白质来进行，例如涉及γ-H2A.X、53BP1或RAD51。我们最近开发了“The Focinator”，作为一种用于细胞核和DNA损伤病灶自动定量和定性分析的可靠且快速的工具。由于具有图形界面和更多功能，这款经过改进的软件现在更加用户友好。因此，我们纳入了基于R脚本的模式，用于自动图像打开、文件命名、进度监测和错误报告。因此，在初始参数定义后，评估不再需要操作员在场。此外，Focinator v2-0现在能够对四个通道进行多通道分析，并通过比较多达三个病灶通道来评估蛋白质-蛋白质共定位。这使得例如能够对特定细胞周期阶段的细胞中的病灶进行定量分析。
