Doxorubicin is overlapping in FL-2 channel and the profile looks indistinct. Has anyone faced such problem? any solution?
Doxorubicin is fluorescent in the orange/red channel. No way to double stain with PI even after extensive washing, since the drug is retained in the nucleus in a dose dependent way.
Not sure if you want to make cell cycle analysis or live/dead assay. What I am writing it would apply only to the second case.
The only solution is to use alternative dyes. The SYTOX green DNA dye should work since it is a green emitting dye with excitation in the blue, so with the same laser used for PI. You should not get spillover of the doxorubicin fluorescence in the green channel.
Hope this helps.
Depending on the machine you are working on, you could also try DAPI, as it also does not permeate live cells.
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