DiOC6(3) strange staining - please, can a specialist of this dye help me? (Nov/17/2005 )

hi,

i have incubated my Jurkat cells with a compound for 16h before a DiOC6(3) staining of apoptotic cells
i used negative and positive controls and obtained for the first one a positively marked population on FL1, for the second one a negatively marked population with very clean histograms...

the problem is that with my compound i obtained a doubled peak in the positively marked population (non-apoptotic) (and a single peak in the negatively marked population) and i can't explain this phenomenum!

since the dye act on mitochondria, can we say that there is a difference in the mitochondrial membrane potential in the non-apoptotic cells?

thanks a lot

Seb_

Hi, i am not a specialist but i use dioc 6 so maybe i can help you..... i think that you are right when you say "there is a difference in the mitochondrial membrane potential in the non-apoptotic cells"... i have observed that cells that are committed to death show an increase in the fluorescence when stained with DiOC(6).... so maybe the "more positive" peak are cells that start to be apoptotic...

Good luck