[Histonet] question about inmuno

[Jose Luis Palazon Fernandez]

Dear List-Members

First I want to thank the people who kindly answered to my question about acid-resistant haematoxylin. I have a question about inmunohistochemistry. I have to make a inmuno to detect ATPase but then I would like to make another staining like PAS or H&E. Could these "counterstainins" affect the inmuno reaction or even cuould the inmuno response be lost or masked?. I want to distinguish to kinds of cells, both of then are positive to PAS but only one is positive to ATPase, I would like to show the two in the same picture. Any help or comentary would be appreciated

thanks in advance

José Luis




El dia 20/09/2004 21:13 usted envio el siguiente mensaje:
>Date: 20 de Septiembre de 2004 21:13:32
>From: "Gudrun Lang" <gu.lang <@t> gmx.at> 
>Subject: Re: [Histonet] hematoxylin 
>To: jluis.palazon <@t> icman.csic.es
>
> I think the usual substitute here is Weigert's Hematoxylin. It stains the
> nuclei black. We use it at the beginning of Trichrom-stains.
> staining: 5 min (or more), differentiation with 0,5% HCl-Alk. (obtional),
> blueing 5 min tapwater.
> 
> working solution:
> Gieson A + Gieson B 1:1 (stable for one week, older solutions stain rather
> brownish than black)
> 
> Gieson A:
> 10 g hematoxylin + 1000 ml 96% Alk. (allow to stand for one week)
> 
> Gieson B:
> 40 ml 29% Ferric-chlorid
> 980 ml Aqua dest
> 10 ml 25% HCl
> 
> greetings
> Gudrun
> 
> 
> ----- Original Message ----- 
> From: "Jose Luis Palazon Fernandez" <jluis.palazon <@t> icman.csic.es>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Monday, September 20, 2004 8:25 PM
> Subject: [Histonet] hematoxylin
> 
> 
> Dear List-members
> 
> greetings. I would like to know if there is a substitute for the "celestine
> blue/hematoxylin" staining of nucleus that can resist a posterior acidic
> counterstain. I need to apply a protocol that uses this nuclear staining
> method but I dont have celestine blue and the company that suplies it says
> that it would last 2 months to supply the stain
> 
> Thanks in advance
> 
> José Luis
> 
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Universidad de Oriente-Isla Margarita-Venezuela
actualmente en: Instituto de Ciencias Marinas de Andalucia
Puerto Real, Cádiz, España.
email: jluis.palazon <@t> icman.csic.es