Re: Viability / nuclear stain

From: Deborah Berglund (umbbd@gemini.oscs.montana.edu)
Date: Tue Mar 09 1999 - 14:22:51 EST


PI depends on membrane integrity for exclusion.  If PI doesn't work then
any other fluorochrome dye that also is dependent on membrane integrity
will probably do the same thing.  You might try FDA, labels viable cells
due to nonspecific esterase activity, but it might leak out of the live
cells if they are permeable.  Your best bet is to improve your preparation
in order not to cause the live cells to become permeable.

Deb Berglund

On Mon, 8 Mar 1999, Sue DeMaggio wrote:

> 
> 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>http://mamba.bio.uci.e
> du/~suedemag/OBCore/index.htm



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