Need a simple procedure for PI/Cell surface Staining

From: Bill Justice (bjustice@kumc.edu)
Date: Fri Mar 09 2001 - 15:57:59 EST 

I am sure this is a very simplistic question, but that's good because I am a very
simple person! (grin)

I have an investigator that is staining mice cells for CD3 (biotin-strepavidin),
CD4, and CD8. In addition the investigator wants to see the cell cycle using PI for
various populations.

I know that I will need to Aldehyde fix the cells first with the cell surface, but need
to know a simple procedure from that point to permeablize the cells while leaving the
cell membrane intact to stain with the PI.

I am sure it will involve using VERY COLD %70 EtOH on ice for a specific amount of time,
then washes and then introduction of the PI Stock reagent less detergent.

Does anyone have a specific procedure to accommodate this request I could use?

Thanks in advance.




Bill Justice
Laboratory Information Systems
KU Medical Center

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