[Histonet] RE: MOHS IPs

[Barbara Tibbs]

Hello Karen,

Back in the old days of doing IHC manually on mostly fresh, frozen tissue I would immediately place the slide with the frozen section into a coplin jar of acetone to fix it.  Keep the coplin jar of acetone in the cryostat.   Leave it in the acetone for only 10 minutes.  Try diluting the Protease 2 - 1:1 to make it gentler so it doesn't "eat" the tissue.   This technique worked well when I was doing ER/PR on frozen breast tumors.  Not sure it will work with skin but it's worth a try.

Barbara S. Tibbs
Histology Supervisor
Accurate Diagnostic Labs
South Plainfield, NJ
barbara.tibbs <@t> accuratediagnosticlabs.com
732-839-3374
Cell: 610-809-6508


________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu <histonet-bounces <@t> lists.utsouthwestern.edu> on behalf of Bauer, Karen L. <Bauer.Karen <@t> mayo.edu>
Sent: Tuesday, October 01, 2013 5:22 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] MOHS IPs

Hi all,

We are in the process of validating some antibodies in our MOHS lab.  The Melan A (Mart 1) antibody is working well, but it could be darker.  We will be increasing the Ab incubation time to see if that will help.

As for the Pan Keratin, we cannot get it to work at all.  We use Protease 2 on our Ultra stainer for FFPE tissues in Histology, but this stain in MOHS is placed on fresh, unfixed tissue... by a manual drop method.  Any time we've tried to use an enzyme retrieval, the tissue looks eaten up... even if we incubate if for a minute.

Since the tissue is not fixed, I figured that no retrieval step would be needed, but the Pan Keratin refuses to work with or without retrieval.

For those of you in MOHS labs that are using a manual staining method for Melan A and Pan Keratin, would you be willing to share your protocol with us?

Thanks so much!!

Karen

Karen L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology | MOHS Lab Supervisor | Dermatology | Phone: 715-838-3205 | bauer.karen <@t> mayo.edu<mailto:bauer.karen <@t> mayo.edu> | Mayo Clinic Health System | 1221 Whipple Street | Eau Claire, WI 54702 | mayoclinichealthsystem.org<http://www.mayoclinichealthsystem.org/>


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