I’d like to quantify postively stained (intracellular Immunofluorescence) cells. So I identfied nuclei by DAPI staining and identfied them by using the IDENTIFY_PRIMARY_OBJECTS.
I used IDENTIFY_SECONDARY_OBJECTS to identify cell boundaries (stained with phalloidin labelled with TxRed).
Now I’d like to identify positive singals of my specific staining (Green channel) and relate them to the identified cells (DAPI+ Phalloidin). Unfortunately, IDENTIFY_TERTIARY_OBEJECTs didn’t work out. When I use the IDENTIFY_PRIMARY_OBJECTS module (now for the green channel, my specific staining respectively) Cell profiler sucessfully identifies valid signals. How can I now relate the identified objects to the cells and count them?
hopefully anyone can help me.
Thanks a lot