[Histonet] blue d'alcian or whole PAS after ISH

[John Kiernan]

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 <@t> uwo.ca
   http://publish.uwo.ca/~jkiernan/
___________________________

Nancy.Walker <@t> 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 <@t> sanofi-synthelabo.com
> tel : (33)561004179  fax :(33)561004001
> 
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