RE: Viability / nuclear stain

From: Peter Lopez (Peterl@Cytomation.com)
Date: Tue Mar 09 1999 - 12:47:56 EST


Hi Sue,
	You can use HO33258 (if you have a UV laser), if you keep the
cells on ice, and the histogram 
distribution will be similar to what you'd expect using PI. You can look
at the cells (kept on ice)
within a half hour and have excellent (1-2 log) differentiation between
live and dead.
I presented a poster on this at ISAC XIX  directed towards users of arc
lamp single detector
instruments, but it will work fine in a multilaser set-up.
	Simply add HO33258 to an iced cell suspension at 1ug/ml final
concentration. Analysis
can be immediate after addition of dye.

Peter
	

> ----------
> From: 	Sue DeMaggio[SMTP:suedemag@uci.edu]
> Sent: 	Monday, March 08, 1999 12:50 PM
> To: 	Cytometry Mailing List
> Subject: 	Viability / nuclear stain
> 
> Has anyone used anything other than PI to do viability staining on the
> FLOW?  7-AAD?  
> 
> 
> I have a researcher using PI and AO and they co-stain ALL the cells,
> virtually 100%, in what they believe to be live samples  of
> Chondrocytes.  Any hints?  Does the collagenase used to break up the
> matrix of the cartilage cause permeabilization?  I'm kind of at a loss
> to explain to him what is happening.  Any better dye for this
> application?
> 
> Thanks in advance! 
> Susan DeMaggio, MS BSMT(ASCP)QCym 
> Resource Manager 
> Optical Biology Shared Resource 
> University of California, Irvine 
> Irvine, CA  92697-2275 
> phone   949-824-4110 
> fax     949-824-3571 
> 
> http://mamba.bio.uci.edu/~suedemag/OBCore/index.htm  
> 



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