Quantitative image analysis of gamma-H2AX foci induced by ionizing radiation applying open source programs.

Jorge Ernesto González, Manual Lee, Joan Francesc Barquinero, Marco Valente, Sandrine Roch-Lefèvre, Omar García
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Abstract

Objective: To test a CellProfiler pipeline for automated counting and characterization of gamma-H2AX foci in color images of human cultured cells.

Study design: A431 cells were irradiated and stained for gamma-H2AX foci detection. Sets of color images were analyzed visually, and findings were compared with those using FociCounter and CellProfiler software.

Results: The CellProfiler pipeline includes some proprieties not present in FociCounter, such as the automatic detection of nuclei, the detection of touching nuclei and the rejection of nuclei that touch the border of the image. The time required for manual operation is associated with the number of images analyzed visually or by FociCounter but not for the CellProfiler program. CellProfiler reduced manual operation time and is about 4 times faster than semiautomatic detection using FociCounter and 10 times faster than visual counting.

Conclusion: We conclude that CellProfiler and FociCounter are reliable tools for measuring gamma-H2AX foci. However, CellProfiler overcomes the limitations of the FociCounter program and is able to detect nuclei automatically, saving considerable manual operation.

应用开源程序定量分析电离辐射诱导的γ - h2ax聚焦。
目的:测试用于人培养细胞彩色图像中γ - h2ax病灶自动计数和表征的CellProfiler流水线。研究设计:A431细胞辐照染色,进行γ - h2ax病灶检测。对彩色图像集进行视觉分析,并将结果与使用FociCounter和CellProfiler软件的结果进行比较。结果:CellProfiler流水线包含了FociCounter所不具备的一些特性,如自动检测细胞核、检测接触细胞核和拒绝接触图像边界的细胞核。手动操作所需的时间与视觉或FociCounter分析的图像数量有关,但与CellProfiler程序无关。CellProfiler减少了人工操作时间,比使用FociCounter的半自动检测快4倍,比视觉计数快10倍。结论:我们得出CellProfiler和FociCounter是测量γ - h2ax焦点的可靠工具。然而,CellProfiler克服了FociCounter程序的局限性,能够自动检测细胞核,节省了大量的人工操作。
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