Re[2]: Apoptosis with PI stain
tom_frey@bdis.com
Fri, 07 Jun 96 17:10:32
The original posting requested measurement of apoptosis in live cells with PI.
This is usually done by incubating with something like 5 ug/ml for 10-15 minutes
and hoping the apoptotic cells have gotten brighter than non-apoptotic but not
as bright as dead cells. This has been reported for almost all of the dead cell
stains (ethidium bromide, 7-AAD, TO-PRO-3). I would use 7-AAD for this if I was
limited to these choices.
I personally prefer to use LDS-751 (I'm prejudiced) or fluorescein diacteate.
These reagents have less time dependence (note they stain apoptotic DIMMER, not
brighter), and have been very reliable in my hand. FDA has the advantage of
also indicating cells that have gone on to "secondary necrosis" or whatever you
prefer to call the result of the apoptotic process. FDA has the drawback of
using up your fluorescein channel.
Tom_Frey@bdis.com
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