Please help me to make a pipeline


I took lots of imagines for mice spinal cord, and need to count the microglia. The cells were stained with antibody (red) and DAPI (blue). We need to count the microglia with both red and blue. As I am new for CellProfiler and my boss wants the data ASAP, there is no time to learn from the beginning. Would you please help me to make a pipeline?
I attached the imagin here.
Many thanks.


First, BEFORE POSTING - Please read this

Basically, a pipeline would be:
(1) LoadImages (DAPI and Ab)
(2) ColorToGray
(3) IdentifyPrimaryObjects (DAPI).
?? Is the Ab in nuclear compartment, as well as the cytoplasm?
(4) IdentifySecondaryObjects, using the nucleus as the seed, and the Ab channel to “grow” into cytoplasm (try “Propagate”)
(5) MeasureObjectIntensity - measure Ab in either nuclear and/or cytoplasm
(6) ExportToSpreadsheet.

We have example pipelines here:
These cells are fairly confluent, so don’t expect great segmentation. You can post your pipeline here and we can comment on it.

Good luck,