Creating a module to measure the intensity of nuclear stain

Hi,

I am new to using CellProfiler and needed some help designing a module to assess the nuclear pH2AX stain in cells from images taken at 20X.

So far I’ve designed a pipeline that looks like this:

Load images
Colortogray (both nuclei (blue) and pH2AX (red))
Identify Primary Objects, nuclei
Identify Primary Objects, pH2AX
MeasureObjectIntensity, measure pH2AX from pH2AXimage
Relate Objects, Input child object= pH2AX, Input parent objects=Nuclei and check YES to calculate per parent means for all child measurements
Display data online, display pH2AX mean instensity
and ExportToSpreadsheet

However, I am unable to see the measurement for per parent mean of child measurements.
Where does this show up?
Also, is there a better pipeline that I could use to assess a nuclear stain using dapi for nuclei parent reference?

I’ve attached example images for reference.

Thanks for your help.



[quote=“nspardy”]However, I am unable to see the measurement for per parent mean of child measurements.
Where does this show up?[/quote]

The per-parent means should show up as columns in the nuclei spreadsheet.

I’ve attached a pipeline which should help get you started. I have the following recommendations re: image acquisition:

  • You have some saturation in your DAPI image which can affect nuclei detection. You may want to dial down the exposure on this channel a bit.
  • As it stands, I need to have extra modules to convert your color images into grayscale. You may want to save your raw images as grayscale upon acquisition to save yourself some time.

Regards,
-Mark
2012_10_01.cp (11.8 KB)