Hi Every one, I want to do mitosox live cell imaging for LX-2 cells. I tried once before, I could nt see any signals of mitosox. I stained cells with 5um of Mitosox dye for 15 minutes at 37 0C and then washed three times with warm buffer. After that I stained cells with Hoechst dye for 10 minutes and then washed three times with wash buffer (PBS+FBS 48ml +2 ml).

I think i washed away all Mitosox. Please let me know what could be the possible reason and what I should try next time.

Thank you very much