Hello everybody,

I need your help concerning the coloration of my liver sections with Bodipy 493/503. Indeed, I would like to quantify the steatosis (to make a score of the NAFLD) with the size of the lipid droplets. The bodipy seems to me to be a good initiative. But I have carried out several tests, and I have the impression that I’m losing information. My lipid droplets aren’t really circular and other areas look "empty". I have tried fixing my liver sections with PFA 4% before and after staining, but have no change. I’m not really surprised since when I collect my tissue, it’s directly put in a PFA 4% solution for 48 hours before including it in an OCT medium.

Do I alter the tissue during the treatment? Or is it during coloring? Do you have any idea where the problem may be coming from?

For information, I wash my slides 3 times 5 minutes in 1X PBS. Then I apply the Bodipy for 30 or 60 minutes (concentration: 50 µg / mL) before washing 3 times 10 minutes in PBS. I then mount my blades.

Thank you for your help or suggestions.