[Histonet] RE: MOHS IPs

[Kim Donadio]

What detection kit are you using? 

Sent from my iPhone

On Oct 2, 2013, at 10:47 AM, "Bauer, Karen L." <Bauer.Karen <@t> mayo.edu> wrote:

> Thanks for the reply,
> 
> We are pre-fixing in a formalin substitute, but we'll give the acetone a try.  I have diluted the protease, but have not gotten any good results.  We'll keep trying with that as well.
> 
> We've tried multiple incubation times for the cytokeratin... From 5 minutes all the way up to 30 minutes... With no luck.
> 
> We are using an enhanced polymer DAB.
> 
> Thanks for the suggestions!  I greatly appreciate it!  :)
> 
> Karen
> 
> Karen L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology | MOHS Lab Supervisor | Dermatology | Phone: 715-838-3205 | bauer.karen <@t> mayo.edu | Mayo Clinic Health System | 1221 Whipple Street | Eau Claire, WI 54702 | mayoclinichealthsystem.org
> 
> 
> 
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of pruegg <@t> ihctech.net
> Sent: Wednesday, October 02, 2013 9:36 AM
> To: Barbara Tibbs; Bauer, Karen L.; 'histonet <@t> lists.utsouthwestern.edu'
> Subject: RE: [Histonet] RE: MOHS IPs
> 
> 
>   I  agree with Barbara try fixing in cold acetone/ethanol mix even =
>   just  for  a  couple  of minutes then go directly to buffer do not dry
>   after=x,  I  would dilute the protease 1:4 with buffer (PBS or TBS
>   what  ever  u  u=) and do it for a short time.  Enzyme digestion on
>   frozen  sections  i=ricky  but  fixing a little and diluting the
>   enzyme  should  help  prevent  ov=  digestion.   How  long  are  u
>   incubating  the  primary?   Are  u usin=n enhanced labeled polymer
>   detection?  HRP/Dab or AEC or Alk.P/fast=d?
> 
> 
> 
> 
> 
> 
> 
> 
> -------- Original Message --------
> 
> Subject: [Histonet] RE: MOHS =s
> 
> From: Barbara Tibbs <[1]barbara.tibbs <@t> accuratediagnosticlabs.com>
> 
> D=e: Wed, October 02, 2013 5:54 am
> 
> To: "Bauer, Karen L." <[2]Bauer.Karen <@t> mayo.edu>,
> 
> "'histonet <@t> lists.utsouthwest=n.edu'"
> 
> <[3]histonet <@t> lists.utsouthwestern.edu>
> 
> 
> 
> Hello Karen,
>   =A
> 
> Back  in the old days of doing IHC manually on mostly fresh, froze   n  tissue  I would immediately place the slide with the frozen section
>   into  a=plin  jar  of  acetone  to  fix it. Keep the coplin jar of
>   acetone  in  the  cr=stat.  Leave  it  in  the acetone for only 10
>   minutes.  Try diluting the Pr=ease 2 - 1:1 to make it gentler so it
>   doesn't  "eat"  the  tissue.  This  te=nique worked well when I was
>   doing ER/PR on frozen breast tumors. Not su= it will work with skin
>   but it's worth a try.
> 
> 
> 
> Barbara S. Tib=
> 
> Histology Supervisor
> 
> Accurate Diagnostic Labs
> 
> South Pl=nfield, NJ
> 
> [4]barbara.tibbs <@t> accuratediagnosticlabs.com
> 
> 732-839-3374
> 
> C=l: 610-809-6508
> 
> 
> 
> 
> 
> _____________________________________=_
> 
> From:                  [5]histonet-bounces <@t> lists.utsouthwestern.edu
>   <[6]histonet-bounces <@t> lists.utsouthwestern=du>  on behalf of Bauer,
>   Karen L. <[7]Bauer.Karen <@t> mayo.edu>
> 
> Sent: Tuesday, October 01, 20 5:22 PM
> 
> To: '[8]=stonet <@t> lists.utsouthwestern.edu'
> 
> Subject: [Histonet] MOHS IPs
> 
> 
> Hi all,
> 
> 
> 
> We  are  in the process of validating some a=ibodies in our MOHS
>   lab.  The Melan A (Mart 1) antibody is working well, =t it could be
>   darker.  We will be increasing the Ab incubation time to se=f that
>   will help.
> 
> 
> 
> As  for  the Pan Keratin, we cannot get it = work at all. We use
>   Protease  2  on  our Ultra stainer for FFPE tissues in=stology, but
>   this  stain in MOHS is placed on fresh, unfixed tissue... by=anual
>   drop  method.  Any  time we've tried to use an enzyme retrieval, th=
>   tissue looks eaten up... even if we incubate if for a minute.
> 
> 
>   =ASince  the tissue is not fixed, I figured that no retrieval step
>   would  be=eded, but the Pan Keratin refuses to work with or without
>   retrieval.
>   =A
> 
> For  those of you in MOHS labs that are using a manual staining me   thod  for  Melan A and Pan Keratin, would you be willing to share your
>   protoc= with us?
> 
> 
> 
> Thanks so much!!
> 
> 
> 
> Karen
> 
> 
> 
> K=en L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology |
>   MOHS   Lab   S=ervisor  |  Dermatology  |  Phone:  715-838-3205  |
>   [9]bauer.karen <@t> mayo.edu<[10]mailto:bauer.karen <@t> mayo.edu> | Mayo Clinic
>   Health  System  |  122=hipple  Street  |  Eau Claire, WI 54702 |
>   [11]mayoclinichealthsystem.org<http://www.[12]mayoclinichealthsystem.o
>   rg/>
> 
> 
>   =A
> 
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> References
> 
>   1. 3D"mailto:barbara.tibbs <@t> accurated   2. ="mailto:Bauer.Karen <@t> mayo.edu"
>   3. 3D"mailto:histonet <@t> lists.utsouthwestern.edu   4. 3D"mailto:barbara.tibbs <@t> accuratediagnosticlabs.c   5. 3D"mailto:histonet-bounces <@t> lists.utsouthwestern.edu   6. 3D"mailto:histo   7. 3D"mailto:Bauer.Karen   8. 3D"mailto:histonet <@t> lists.utsouthwestern.edu"
>   9. 3D"mailto:bauer.kar  10. 3D"mailto:bauer.karen <@t> mayo  11. 3D"http://mayoclinichealt=
>  12. 3D"http:/  13. 3D"mailto:Histonet <@t> lists.utsouthwestern.edu"
>  14. 3D"http://lists.utsouthweste=
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