Automated gamma-H2AX focus scoring method for human lymphocytes after ionizing radiation exposure
Automated gamma-H2AX focus scoring method for human lymphocytes after ionizing radiation exposure
复制标题
DOI：
10.1016/j.radmeas.2011.05.012
复制
发表时间：
2011-09-01
影响因子：
2
通讯作者：
Roch-Lefevre, S.
中科院分区：
文献类型：
--
作者：
Valente, M.;Voisin, P.;Roch-Lefevre, S.
本研究的目的是开发一种基于显微镜的人类淋巴细胞病灶定量方案，能够为辐射敏感性测定提供有用的数据。将人外周血暴露于伽马射线，并用荧光染料偶联的抗伽马-H2AX（丝氨酸 139 的组蛋白 2AX 磷酸化）抗体对分离的淋巴细胞进行染色。自动采集显微镜载玻片，并对所得图像进行 3 种焦点评分方法：手动、半自动和全自动。所有评分方法都足够灵敏，能够检测至少 0.05 Gy 的辐射，且实验之间的差异较小。对于较高的收盘价，两种自动方法往往检测到的焦点少于手动评分，但仍获得线性相关性（最低 r(2) > 0.971）。与对图像进行手动评分相比，自动化方法至少快 5 倍，并且所需的操作员干预最少。我们可以得出结论，我们的方法能够在不到 6 小时的时间内获得血液样本的病灶评分。除了焦点评分之外，所使用的程序还执行一些具有潜在生物学重要性的细胞和焦点测量。 (C) 2011 Elsevier Ltd. 保留所有权利。
The purpose of this study was to develop a microscopy-based foci quantification protocol in human lymphocyte capable of supplying useful data for radiation sensitivity assays. Human peripheral blood was exposed to gamma-rays and isolated lymphocytes were stained with fluorochrome-coupled anti-gamma-H2AX (Histone 2AX phosphorylation of serine 139) antibodies. Microscopy slides were automatically acquired and the resulting images were subjected to 3 focus scoring methods: manual, semi-automated and fully automated. All scoring methods were sufficiently sensitive to detect an irradiation of at least 0.05 Gy with low variation between experiments. For higher closes, both automated approaches tend to detect fewer foci than manual scoring but still obtaining a linear correlation (lowest r(2) > 0.971). Compared with manual scoring on the images, the automated approaches are at least 5 times faster with minimum operator intervention needed. We can conclude that our method is able to obtain the foci score of a blood sample in less than 6 h. In addition to the foci score the programs used perform several cell and foci measurements of potential biological importance. (C) 2011 Elsevier Ltd. All rights reserved.