Hi everyone!

We're using ER-Tracker™ Red (BODIPY® TR Glibenclamide, Invitrogen) to visualize the endoplasmic reticulum (ER) and ER stress in primary hepatocytes treated with tunicamycin (5 µg/mL for 6 hours). However, we're having trouble getting a clean signal. Instead of a clear ER pattern, we see a diffuse red "haze" over the cells, which obscures the specific staining when we image in the AF584 (Alexa Fluor 584) channel.

Has anyone successfully used this dye in primary hepatocytes or similar cells under ER stress conditions? Do you have any tips or protocol modifications that helped improve the signal quality?

Thanks in advance for your help!