CyChrome Hoechst question

From: Andrew Oberyszyn (oberyszyn.2@osu.edu)
Date: Tue Oct 29 2002 - 15:43:43 EST 




Sorry to bother everybody so soon with another question but I came across
an interesting and odd phenomenon which I can't explain.  

I have a user who is doing 4 color staining (FITC, PE, CyChrome,
Hoechst).  She hasn't been able to get good Hoechst CV's with live
Bone Marrow cells so she's gone to PF and EtOH fixation.

The problem occurs when she used room temp EtOH instead of ice cold
EtOH.  Under this condition, the Cychrome (B220) staining was
lost.  

Does anybody have an explanation as to why RT EtOH would have this effect
and not ice cold EtOH?


Just curious!

Thanks for any help in advance!

Andy


P.S.  Neither the FITC or PE staining was affected.



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