Re: strange request...

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From:denise M m Long-Woodward <>

It has been my experience with skin cell cultures that the
collagen/fibrin gel doesn't stain with either H or E.
Why not process just one gel (without cell cultures) to verify?
Denise Long Woodward

On Mon, 24 Apr 2000 16:45:22 -0400 "Johnson, Jennifer(Hist)"
<> writes:
> Dear Histonetters,
> 	An investigator came to me today with a strange request.  
> She is
> growing small pieces of mouse aortas in a collagen and fibrin gel.  
> She
> would like to be able to stain the aorta and the cells growing from 
> it,
> without staining the gel.  She wants to perform the stain on the 
> whole 1-2
> mm thick plug without sectioning it in order to try and count the 
> number of
> cells that have grown off of the aorta.  I have suggested that 
> processing
> into paraffin may give her more success, but she is really looking 
> for a
> quick and dirty instant answer.  (Doesn't this sound familiar?)   
> 	Since, ideally, the method chosen should be relatively 
> simple, I am
> going to first try a nuclear fast red, or maybe a hematoxylin 
> (without
> eosin), or even a DAPI stain.  My thinking is to stain the nuclei, 
> and avoid
> cytoplasm (I think that in these cells the cytoplasm and aorta 
> itself will
> have collagen in them).  
> 	My second idea is to try a fluorescently labeled S-100 
> antibody,
> (using the most abbreviated protocol I can find!) to label the cells 
> only.
> 	But I am having a lot of trouble finding any other methods.  
> Most
> books are organized by what you want to stain, not by what you are 
> trying to
> avoid staining!   If anyone has had any experience with this sort of
> staining, or has any ideas, I would appreciate hearing from you!  
> Thanks,
> Jennifer Johnson

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