I’m trying to use the ClassifyObjectsbyTwoMeasurements module to quantify the number of nuclei with both FITC and TRITC label, either FITC or TRITC and without any label.
The module works fine provided there are measurements below the two threshold values, however if for example all the intensity measurements in the FITC channel are above the threshold (eg all nuclei FITC positive) the following error message pops up and the pipeline stops. Is there something I’m doing wrong here or is that a bug in the module, and if it is not a bug how else could I do that counting ?
There was a problem running the analysis module ClassifyObjectsByTwoMeasurements which is number 18.
Image processing was canceled in the ClassifyObjectsByTwoMeasurements module because the threshold 0.04 you selected for the first set of data is invalid. The threshold should be a number in between the minimum and maximum measurements of the data, which are 0.043171 and 0.054518, respectively.
ClassifyObjectsByTwoMeasurements in C:\Program Files\CellProfiler_105811\CompiledCellProfiler\CellProfiler_mcr\Modules\ClassifyObjectsByTwoMeasurements.m (199)
AnalyzeImagesButton_Callback in C:\Program Files\CellProfiler_105811\CompiledCellProfiler\CellProfiler_mcr\CellProfiler\CellProfiler.m (10424)
gui_mainfcn in C:\Program Files\CellProfiler_105811\CompiledCellProfiler\CellProfiler_mcr\CellProfiler\CellProfiler.m (12164)
CellProfiler in C:\Program Files\CellProfiler_105811\CompiledCellProfiler\CellProfiler_mcr\CellProfiler\CellProfiler.m (57)