[Histonet] question about inmuno

[SMITH,REBEKAH FELICIA]

I would think that it would depend highly on which chromagen 
you're using for your IHC. I have done IHC and H&E on the same 
slide, but it does make it a lot harder to see where your 
immunostaining is(especially since I was using brown DAB). My 
immunostaining was still there, btw, it was just really hard to 
see. Perhaps try  to either get or make the DAB that turns black 
rather than brown (I think thats the nickel DAB) or use a 
chromagen thats a completely different color outside of the 
brownish, pinkish or bluish range. (I'm not sure what other 
chromogens there are since I've only used AEC and DAB but I know 
Vector makes some others.)
                                          Rebekah Smith
On Thu Sep 23 12:39:35 EDT 2004, Jose Luis Palazon Fernandez 
<jluis.palazon <@t> icman.csic.es> wrote:

> 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
> 
> 
> 
> Jos? 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
> 
>> 
> 
>> Jos? Luis
> 
>> 
> 
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> 
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>> 
> 
> 
> 
> 
> 
> 
> 
> Universidad de Oriente-Isla Margarita-Venezuela
> 
> actualmente en: Instituto de Ciencias Marinas de Andalucia
> 
> Puerto Real, C?diz, Espa?a.
> 
> email: jluis.palazon <@t> icman.csic.es
> 
> 
> 
> 
> 
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> 



--
SMITH,REBEKAH FELICIA
"You are a child of the universe, no less than the trees and the 
stars
You have a right to be here and whether or not it is clear to you, 
no doubt the universe is unfolding as it should. Therefore be at 
peace with G-d, whatever you conceive Him to be. And whatever your 
labors and aspirations,in the noisy confusion of life, keep peace 
in your soul.-Max Ehrmann,"Desiderata"