Nuclei are not always in center of cell in confocal images. So when secondary object identfication is conducted, it gives the incorrect locations for even obvious cell borders. Any way I can isolate the border and identify cells without using the nuclei?
In IdentifySecondaryObjects, you may want to use "Watershed - Image" instead of "Propagation", with Adaptive - Otsu - Three classes.
It may not be perfect, but can still help.