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----- From: [email protected] [mailto:[email protected]]on Behalf Of [email protected] Sent: Saturday, July 25, 2009 1:01 PM To: [email protected] Subject: Histonet Digest, Vol 68, Issue 32 Send Histonet mailing list submissions to [email protected] To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to [email protected] You can reach the person managing the list at [email protected] When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." 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) ---------------------------------------------------------------------- Message: 1 Date: Fri, 24 Jul 2009 13:01:17 -0400 From: "Diana McCaig" <[email protected]> Subject: [Histonet] IMMUNO STAINING--DECOLORIZING To: <[email protected]> Message-ID: <[email protected]> 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 ------------------------------ Message: 2 Date: Fri, 24 Jul 2009 13:07:26 -0400 From: "McMahon, Loralee A" <[email protected]> Subject: RE: [Histonet] IMMUNO STAINING--DECOLORIZING To: Diana McCaig <[email protected]>, <[email protected]> Message-ID: <2cf6f6b05263ea4ebab07781b51e5db002c94...@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: [email protected] on behalf of Diana McCaig Sent: Fri 7/24/2009 1:01 PM To: [email protected] 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 _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Fri, 24 Jul 2009 20:51:39 -0500 From: "Joe Nocito" <[email protected]> Subject: Re: [Histonet] GMS-Fungus on nails To: "jstaruk" <[email protected]>, "'Clare Thornton'" <[email protected]>, <[email protected]> Message-ID: <ec45792f0876405e9d866c6ebe51b...@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" <[email protected]> To: "'Clare Thornton'" <[email protected]>; <[email protected]> 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: [email protected] > [mailto:[email protected]] On Behalf Of Clare > Thornton > Sent: Friday, July 24, 2009 11:31 AM > To: '[email protected]' > 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 > [email protected] > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 68, Issue 32 **************************************** _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
