[Histonet] RE: Histonet Digest, Vol 68, Issue 32

Mon Jul 27 10:45:05 CDT 2009

We have taken single IHC stains back as far as heat retrieval with excellent results. We have never done this with a cocktail...(but hey, maybe you have a technical paper in the making). The antigens most likely are still there....go back through the obvious steps, and restain using the same protocol. Don't try some other one....you will "muddle" the cells and lose crisp nuclei! Let me know how that works out with a cocktail, if you try it!

-----Original Message-----
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Subject: Histonet Digest, Vol 68, Issue 32

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Today's Topics:

   1. IMMUNO STAINING--DECOLORIZING (Diana McCaig)
   2. RE: IMMUNO STAINING--DECOLORIZING (McMahon, Loralee A)
   3. Re: GMS-Fungus on nails (Joe Nocito)

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Message: 1
Date: Fri, 24 Jul 2009 13:01:17 -0400
From: "Diana McCaig" <dmccaig <@t> ckha.on.ca>
Subject: [Histonet] IMMUNO STAINING--DECOLORIZING
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<DCFD9E6A390E294AAF3A2561CD32E5C40D5775B4 <@t> ckhamail1.ckha.on.ca>
Content-Type: text/plain;	charset="us-ascii"

We had a series of PIN 4 cocktail immuno stains and got a poor reaction.
I am afraid to cut deeper in the block and miss the area of concern.
I have determined the problem was due to the heat bar not being turned
on.

What can I do the slides to reverse the process and restain them.  The
DAB stained good, it is the vulcan fast red that was not reacting.

Diana 

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Message: 2
Date: Fri, 24 Jul 2009 13:07:26 -0400
From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
Subject: RE: [Histonet] IMMUNO STAINING--DECOLORIZING
To: Diana McCaig <dmccaig <@t> ckha.on.ca>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<2CF6F6B05263EA4EBAB07781B51E5DB002C949D5 <@t> e2k3ms1.urmc-sh.rochester.edu>

Content-Type: text/plain; charset="iso-8859-1"

We have had similar problems with the Vulcan Fast Red not staining.  It was suggested by the vendor to buy the fast red in smaller amounts (they actually repackaged the product) so that the chromogen was not being taken out of the fridge and put back in too many times.  Depending on your workflow you may want to aliquot out the chromogen for each run or each couple of runs?  
When we see it starting to fade we use fresh fast red.  That usually takes care of the problem.  
I have never had any luck restaining with the fast red.  And I have tried many times. 

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
University of Rochester 
Department of Pathology

(585) 275-7210

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Diana McCaig
Sent: Fri 7/24/2009 1:01 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IMMUNO STAINING--DECOLORIZING

We had a series of PIN 4 cocktail immuno stains and got a poor reaction.
I am afraid to cut deeper in the block and miss the area of concern.
I have determined the problem was due to the heat bar not being turned
on.

What can I do the slides to reverse the process and restain them.  The
DAB stained good, it is the vulcan fast red that was not reacting.

Diana
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Message: 3
Date: Fri, 24 Jul 2009 20:51:39 -0500
From: "Joe Nocito" <jnocito <@t> satx.rr.com>
Subject: Re: [Histonet] GMS-Fungus on nails
To: "jstaruk" <jstaruk <@t> masshistology.com>,	"'Clare Thornton'"
	<CThornton <@t> dahlchase.com>,	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <EC45792F0876405E9D866C6EBE51BE5D <@t> JoePC>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=original

We soak our nails in 20% sodium hydroxide for at least an hour, rinse in 
running tap water for a few minutes before  we place the cassettes on the 
tissue processor. We use positive charged slides, heat slides in 80 degree 
oven for 20 minutes and perform a PAS/fungus on the slides. The PAS is a lot 
more gentler on the tissue that the GMS. We are the toenail lab for the U.S. 
Air Force ,

Joe "The Toe"

----- Original Message ----- 
From: "jstaruk" <jstaruk <@t> masshistology.com>
To: "'Clare Thornton'" <CThornton <@t> dahlchase.com>; 
<histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, July 24, 2009 10:41 AM
Subject: RE: [Histonet] GMS-Fungus on nails

> Prepare a 10% solution of Titebond II premium wood glue (found in most
> hardware stores).  Dip the slide in the solution just before mounting the
> section on the slide.  Let the slide dry and stain away.
>
> Jim
>
> _______________________
> James E. Staruk HT(ASCP)
> www.masshistology.com
>   www.nehorselabs.com
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Clare
> Thornton
> Sent: Friday, July 24, 2009 11:31 AM
> To: 'histonet <@t> lists.utsouthwestern.edu'
> Subject: [Histonet] GMS-Fungus on nails
>
> We do a GMS for fungus on all nails we receive.  We often have a difficult
> time keeping the nail tissue on the slide.  We've tried baking for long
> periods, pre-treating in formalin, using silane slides, with no luck. 
> Even
> when the nails cut relatively easily we still lose it, and end up running
> several GMS stains before we might get a speck or two of tissue we can 
> look
> at.  We use the Artisan stainer for our specials.  We are really not
> interested in performing the GMS manually, due to volume and turn around
> time restrictions.  Any suggestions?
>
> Clare J. Thornton, HTL(ASCP)
> Assistant Histology Supervisor
> Dahl-Chase Diagnostic Services
> 417 State Street, Suite 540
> Bangor, ME 04401
> cthornton <@t> dahlchase.com
>
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