Identification of myonuclei




I am a new user of ImageJ and CellProfiler. I have images of human muscle sections with four staining channels:

  1. Basal lamina
  2. Nuclei (with Hoechst)
  3. Satellite cells (with pax 7)
  4. Methylation (with 5-MeC)

The ultimate goal is to quantify the intensity of the methylation staining inside the myonuclei and inside the nuclei of the satellite cells.
After reading a lot on ImageJ I was able to already quantify the intensity of the methylation (only the stained 5-MeC that is overlapping nuclei). But now I want to do this specifically for myonuclei and nuclei from satellite cells. Myonuclei are identified based on their location: nuclei with their geometrical centre inside the inner rim of the dystrophin ring (so inside the basal lamina). So easy put: those Hoechst spots that are inside the lamina and not on top of the lamina. Satellite cells can be identified as those myonuclei that show an overlap with the pax 7 signal.

Since there will be around hundred myonuclei within one image and since I have around 500 images, I was hoping there was a way to not do this manually and build a pipeline for this. Are there existing pipelines on this topic that are reliable? Because I am not really good at programming it myself.

Can somebody help me out?



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