Open the options-dialog by right-clicking on the s-button:
#Imagej quantify he staining install#
To install the tool, drag the link MRI_Lipid_Droplets_Tool.ijm to the ImageJ launcher window, save it under macros/toolsets in the ImageJ installation and restart ImageJ. Additional correlative analysis of experimental repeats can be found in Figures S1, S2, S3, S4.The Lipid Droplets Tool helps to segment lipid droplets marked with BODIPY. Regression lines are drawn and the Pearson product moment correlation coefficients ‘r’ is displayed for each set of data. ( C) Correlation analysis of colony intensity percentage fromĪnd the corresponding absorbance data. That were obtained using the colony area percentage and those obtained using the absorbance in ( A). Dots correspond to averages and error bars to the standard deviations of the exact same four wells that were analyzed in ( A) Examples of dose response curves using the optical density from the absorbance measurements of the dye IC 50 = 36.1☖.0 nM (UCN-01) and IC 50 = 6.2☐.5 nM (STS). Were analyzed using a method where the absorption of the crystal violet dye that was washed out from labeled cells is measured. The identical wells that were quantified with ColonyArea in Regression lines are drawn and the Pearson product moment correlation coefficients ‘r’ is displayed for each data set. ( C) Correlation analysis of results obtained using the colony area percentage and results obtained using the colony intensity percentage. Additional independent experimental repeats can be found in Figures S1, S2, S3, S4. Dots correspond to averages and error bars to the standard deviations of measurements from four wells. ( B) Examples of dose response curves using the colony intensity percentage IC 50 = 37.5±5.7 nM (UCN-01) and IC 50 = 16.1☑.4 nM (STS). ( A) Examples of dose response curves using the colony area percentage IC 50 = 35.8±4.5 nM (UCN-01) and IC 50 = 16.4☑.8 nM (STS). Its output parameters were then used to generate dose response curves and determine the half maximal inhibitory concentrations (IC 50) of the compounds.
Image data were analyzed using ColonyArea. We expect that ColonyArea will be of broad utility for cancer biologists, as well as clinical radiation scientists.Ĭolony formation of T98G human glioma cells was studied after treatment with increasing concentrations of the staurosporine derivative UCN-01 or staurosporine (STS).
#Imagej quantify he staining download#
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 IC50 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. 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.
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