Mesenchymal stem cells

I am an MD new to the lab, so I apologize if I am missing something basic. I have been having trouble tracing my cells well. I have tried setting a threshold but haven’t had much success, and when that works it is often for a specific image only and not replicable for other images that may be brighter or darker. Is there a better way? I am using Hoechst stain for my nuclei and cell trackers for cells.
Thanks
David



pipeline for N and C CMFDA.cp (5.6 KB)

Hi David,

Your pipeline is most of the way there, I think, but it appears as though you need some illumination correction on the CellTracker image. I’m attaching a modified version of your pipeline which does this. Also note that the threshold correction factor in IdentifySecondaryObjects may need further adjustment as you continue to test with other images.

Regards,
-Mark
2013_08_23.cp (7.05 KB)

Thank you Dr Bray, that was very helpful.
-David