I want to culture cells after staining with propidium iodide, so there will be PI in the media in which the cells are cultured. Can I do this or will the PI kill the cells? basically I want to stain 150ul of cells and then put 75ul of those cells, with the PI still in the media, into 500ul final culture volume. So there will be a greater than 5x dilution of the PI. Will this be ok or will the residual PI be toxic?
Katja Werner No I don't want to cause any mutations or other changes to the DNA. I was hopeful that since the PI is supposed to be impermeant in living cells I could just leave it in the culture medium diluted 5x and it will not have a deleterious impact on the culture.
I intend to treat the cells, stain them with PI after treatment and flow half of them on the flow cytometer to get an idea of the viability immediately post treatment. The other half I want to put into culture, incubate for ~48 hours and then assess the total number of viable cells from each treatment condition using an ATP assay from Promega.
Its important that the PI remaining in the media from the staining before flow doesn't negatively impact the proliferation of the cells in culture since this would confound with the affect of the treatment.
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