FACS using DAPI for apoptosis analysis - (Jun/03/2013 )
I am treating cancer cells with a drug and want to analyse cancer cell death. The only problem is that the drug auto-fluoresce a lot, so much that I cannot use conventional viable stains like PI or 7AAD (because sometimes spillover cannot be corrected).
I think using DAPI could solve my problem. However, I am not familiar with DAPI. It is often used to determine cell cycle progress, I think, by histology. However, the use of DAPI in FACS would require a much different protocol, I guess (fixation ect.).
I need help, where could I find a good protocol that uses DAPI for the study of apoptosis by FACS ?
With PI you need to fix too, so how do you stain with PI? I don't understand.
Maybe by that he means that he thinks a different kind of fixation might be used...
I've no experience with DAPI in FACS, but did you see this paper ? http://jcb.rupress.org/content/171/3/559.full
However, there are additional alternatives. Try Hoescht 33342 from invitrogen. It fluoresces in the UV range, which I assume DAPI will too, but I know it works by flow. You do have to check that the equipment you are using has the correct filters and laser, though.