RE: strange request...

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From:Cynthia Favara <>
To:"" <>, "'Johnson, Jennifer(Hist)'" <>

	I have stainied tissue in collagen gel and had not problem. might
just want to run the gel through first without tissue. Could be useful.

Cynthia Favara
Rocky Mountain Laboratories
903 S 4th Street
Hamilton, MT 59840
ph: 406-363-9317
FAX: 406-363-9286

> ----------
> From: 	Johnson, Jennifer(Hist)[]
> Sent: 	Monday, April 24, 2000 2:45 PM
> To:
> Subject: 	strange request...
> 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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