[Histonet] question about inmuno

From: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"  

>Subject: Re: [Histonet] hematoxylin 

>To: jluis.palazon@icman.csic.es


> I think the usual substitute here is Weigert's Hematoxylin. It stains the=0D
> nuclei black. We use it at the beginning of Trichrom-stains.

> staining: 5 min (or more), differentiation with 0,5% HCl-Alk. (obtional),=0D
> 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" 

> To: 

> 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=0D
> counterstain. I need to apply a protocol that uses this nuclear staining=0D
> 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@icman.csic.es

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