Re: [Histonet] blue d'alcian or whole PAS after ISH
An alcian blue solution should not damage your
developed and fixed autoradiographic emulsion.
The periodic acid step of PAS might dissolve
the silver grains. When I did autoradiography
(labelled proteins or DNA) I stained the sections
before coating them with the emulsion. In your
tissues that's probably not appropriate because
it might extract the mRNA that you're interested
in.
--
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan@uwo.ca
http://publish.uwo.ca/~jkiernan/
___________________________
Nancy.Walker@sanofi-synthelabo.com wrote:
>
> Help, I need a quick answer don't have time to try to figure things out by
> myself and most of all only have a few slides so I don't want to lose the
> results.
>
> Will Alcian Blue staining alone at pH 4 or Alcian Blue +PAS destroy my
> photographic emulsion coating? I used Alcian Bl + PAS technique long ago
> and it was wonderful for identifying the different compartments of the GI
> tract and identifying the cell types and which were secreting more. Now I
> have an orphan receptor (all we know is it is a putative chloride channel
> activated by chloride) that the mRNA is expressed in the secreting cells of
> the GI tract but in a differential way, I want to see if the mRNA level is
> a function of the level of secretion. or cell types etc. So can I do a bl
> Alcian / PAS or one or the other without destroying my emulsion and theryby
> lose my signal??????????
>
> Has anyone tried to do either of these techniques after microscopic
> photographic emulsion???? or if not can you suggest another technique???
>
> faithfully yours,
> Nancy Walker
> Molecular Biology Scientist
>
> Sanofi-Synthelbo Research
> B.P. 37 Labége Innopole
> 31676 LABEGE CEDEX FRANCE
>
> nancy.walker@sanofi-synthelabo.com
> tel : (33)561004179 fax :(33)561004001
>
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