ColonyArea: an ImageJ plugin to automatically quantify colony formation in clonogenic assays.
ColonyArea: an ImageJ plugin to automatically quantify colony formation in clonogenic assays.
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DOI：
10.1371/journal.pone.0092444
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Abankwa D
中科院分区：
文献类型：
--
作者：
Guzmán C;Bagga M;Kaur A;Westermarck J;Abankwa D
克隆形成或集落形成试验是一种广泛使用的方法，用于研究照射或细胞毒性剂给药后残留的癌细胞集落的数量和大小，并用作这些治疗的抗增殖作用的量度。或者，该测定用于定量癌症相关基因和化学试剂的转化潜力。因此，对于常规实验室使用和并行自动化分析，需要对菌落形成测定进行简化和标准化分析。在这里，我们描述了免费提供的ImageJ插件“ColonyArea”，它是优化的快速和定量分析的焦点形成试验中进行的6至24孔培养皿。ColonyArea处理多孔培养皿的图像数据，在菌落形成定量之前，通过单独分离、同心裁剪和背景校正孔图像。ColonyArea不计算菌落数，而是确定结晶紫染色细胞菌落覆盖面积的百分比，同时考虑染色强度和细胞密度。我们证明，这些参数单独或组合允许两种星形孢菌素，UCN-01和星形孢菌素（STS）对人胶质母细胞瘤细胞（T98 G）的细胞毒性作用的IC 50值的稳健定量。这两种化合物的效力之间的关系与从基于吸光度的方法获得的定量菌落生长和公开数据的关系非常好。ColonyArea ImageJ插件提供了一个简单有效的分析程序，用于定量最常用的细胞测定之一的测定数据。该捆绑包可作为支持信息免费下载。我们预计ColonyArea将为癌症生物学家和临床放射科学家提供广泛的实用性。
The clonogenic or colony formation assay is a widely used method to study the number and size of cancer cell colonies that remain after irradiation or cytotoxic agent administration and serves as a measure for the anti-proliferative effect of these treatments. Alternatively, this assay is used to quantitate the transforming potential of cancer associated genes and chemical agents. Therefore, there is a need for a simplified and standardized analysis of colony formation assays for both routine laboratory use and for parallelized automated analysis. Here we describe the freely available ImageJ-plugin “ColonyArea”, which is optimized for rapid and quantitative analysis of focus formation assays conducted in 6- to 24-well dishes. ColonyArea processes image data of multi-well dishes, by separating, concentrically cropping and background correcting well images individually, before colony formation is quantitated. Instead of counting the number of colonies, ColonyArea determines the percentage of area covered by crystal violet stained cell colonies, also taking the intensity of the staining and therefore cell density into account. We demonstrate that these parameters alone or in combination allow for robust quantification of IC50 values of the cytotoxic effect of two staurosporines, UCN-01 and staurosporine (STS) on human glioblastoma cells (T98G). The relation between the potencies of the two compounds compared very well with that obtained from an absorbance based method to quantify colony growth and to published data. The ColonyArea ImageJ plugin provides a simple and efficient analysis routine to quantitate assay data of one of the most commonly used cellular assays. The bundle is freely available for download as supporting information. We expect that ColonyArea will be of broad utility for cancer biologists, as well as clinical radiation scientists.
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