Relate module

Hi,

Thanks for the great software!! I have a question concerning the relate module. I am using a genetically encode biosensor from an heterozygous population of cells and I want to eliminate cells that don’t express the biosensor before doing any measurements. To do that I have identified, using identify primary object, the cells and the biosensor. I then use the relate module and used the biosensor as the ‘Parent’ and the cells as the ‘Child’ and the relate module keeps only the cells expressing the biosensor ‘Cells with Biosensor’. I would like to use this image (or object kept in the relate module) in the identify ternary object module to substract from the ‘Cells with Biosensor’ the actual staining of the biosensor to give me a ternary object that would be ‘Cytoplasm unoccupied by biosensor’. I could then measure the total area and have an idea of the area occupied in each cells by the biosensor. I have try the filterbyobjectmeasurement but I wasn’t able to get the module to work with the children or parent measurements. I also tried to do it manually in excel with the raw measurements, but it gets pretty confusing in relating each biosensor object to a specific cell.

Thanks a lot for your help
Steve

I think the easiest way to do this might be to use ImageMath to invert the biosensor image, ApplyThreshold to find all the bright pixels in the inverted image (i.e., where the biosensor is not present), and then MaskObjects with the thresholded image to remove those parts of the cytoplasm that have the biosensor.

Thank you so much for your help, it works now. I ended up following what you suggested but I added one more maskobject module. In the first maskobject module I masked the cells using the inverted mask from the tresholded biosensor image and I selected ‘keep’ in 'handling of objects that are partially masked. Then in the second maskobject module I used the masked cells from the previous masked module (only the one with the biosensor staining) and I use again the tresholded biosensor image and did not invert the mask this gave me the cytoplasmic area for all cells expressing the biosensor.
Again many thanks!!