Re: PI staining: reply

J. Paul Robinson (PAUL@flowcyt.cyto.purdue.edu)
Wed, 9 Jul 1997 08:30:51

Reply below Posted for general viewing....
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>Hi-
>
>I hope this is the proper route to post something on the archive. I've
>tracked down several protocols for propidium staining, but I do have one
>question they do not address. Don't you need to centrifuge the stained
>samples before running them through the flow cytometer? I would
>imagine that you still have some PI in solution, and that it would give
>you an awfully high background signal. Any comments?
>
>Thanks. This is a wonderful forum.
>
>Bob Crow
>crow@cbnmr.tamu.edu
>----------------------------------

Bob,
We routinely use PI staining for live/dead discrimination. It does not need
to be washed out prior to running. In fact, PI will leach out if washed. We
add it to a tube, wait a few seconds, and then run the sample on our
FACSCAN. If you are doing PI for DNA where the cells are permeabilized, I
would imagine the same routine applies as PI is reversible in its binding
to DNA. I am sending a copy of this to Paul in hopes he will have it posted
for the group's info.

Good Luck,

Randy Fischer
NIH/NIA

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posted by

J.Paul Robinson, Purdue University Cytometry Labs
Professor of Immunopharmacology
robinson@flowcyt.cyto.purdue.edu PH:765-494 6449 FAX:765-494 0517
web http://www.cyto.purdue.edu