![]() We expect that ColonyArea will be of broad utility for cancer biologists, as well as clinical radiation scientists. The bundle is freely available for download as supporting information. 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 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. We demonstrate that these parameters alone or in combination allow for robust quantification of IC 50 values of the cytotoxic effect of two staurosporines, UCN-01 and staurosporine (STS) on human glioblastoma cells (T98G). 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. ColonyArea processes image data of multi-well dishes, by separating, concentrically cropping and background correcting well images individually, before colony formation is quantitated. 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. 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. Alternatively, this assay is used to quantitate the transforming potential of cancer associated genes and chemical agents. No commercial reproduction, distribution, display or performance rights in this work are provided.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. Cellular assay, imaging, analysis, writing macro: PC. Contributed reagents/materials/analysis tools: PC. The authors have declared that no competing interests exist.Īuthor Contributions: Conceived and designed the experiments: PC. įunding: The authors have no support or funding to report. The IMJ macro, CellProfiler pipelines, and images are currently available from. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability Statement: All relevant data are within the paper and its Supporting Information files. It can fulfill the need to automate colony/cell counting in high-throughput screens, colony forming assays, and cellular assays. In this study, I demonstrate that Cell Colony Edge is superior to other open-source methods, in speed, accuracy and applicability to diverse cellular assays. The ImageJ macro Cell Colony Edge is valuable in counting cells and colonies, and measuring their area, volume, morphology, and intensity. For this purpose, I developed an ImageJ macro Cell Colony Edge and a CellProfiler Pipeline Cell Colony Counting, and compared them to other open-source digital methods and manual counts. The objective of this study was to develop an automated method for quick and reliable counting of cells and colonies from digital images. Due to its subjective and time-intensive nature, manual counting has hindered the adoption of cellular assays such as tumor spheroid formation in high-throughput screens. Use this Persistent URL to link to this item: Ĭounting cells and colonies is an integral part of high-throughput screens and quantitative cellular assays. Other methods of colony counting not tested in paper) Variability in Manual measurement of bacterial colonies) Image Processing steps of IMJ Edge, and selection of parameters) Instructions for downloading and running the macro) ![]() Instructions for manual determination of parameters using ImageJ GUI) Customized versions of Cell_Colony_Edge macro for specific purposes) High-Throughput Method for Automated Colony and Cell Counting by Digital Image Analysis Based on Edge Detection.
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