Re: Viability / nuclear stain

From: Derek Schulze (flow@post.queensu.ca)
Date: Wed Mar 10 1999 - 09:34:53 EST


At 11:50 AM 1999-03-08 -0800, 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?
>Any better dye for this application?

It may not be PI you are seeing.  AO has the unique property of shifting
its emission to red when at a high enough concentration.  This is often how
it is used to visualize the transgolgi (I believe).  It accumulates in the
golgi and after a while turns red.  I guess what I am suggesting is that
you analyze your samples ASAP as the length of time the samples are exposed
to the AO will determine how much red you will see.

Good luck!



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