Re: strange request...
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From: | denise M m Long-Woodward <denisew2@juno.com> |
To: | Jennifer.Johnson@genzyme.com |
Reply-To: | |
Content-Type: | text/plain |
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)"
<Jennifer.Johnson@genzyme.com> 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
> jennifer.johnson@genzyme.com
>
>
>
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