Immunhistochemistry Fibronectin - Fibrosis qunatification?


I search for a method to quantify Fibrosis in heart tissue.The method I found quite useful is: RGB-Stack → Adjust → Threshold

The only problem is that:

  1. Changing the lower threshold doesn’t change the results, I wonder why…(?). It should represent the whole area, which should change if there are black areas for example. I therefore need a method to exclude black areas (which are due to the tissue ripping apart)

  2. I have to manually adjust each threshold, which is different for each picture of course. In my lab they had a macros (doesn’t work anymore), which took the mean of the threshold. Would that make sense?

If anyone has a guide how to do the quantification differently, I would really appreciate that :slight_smile: