[Histonet] decal affecting IHC

[Patsy Ruegg]

I agree with Liz and use 5-10% formic acid for decal for IHC work, on the
other hand if you are trying to demonstrate iron deposits or some enzyme
histochemical stains such as TRAP or Alk.phos you may need to use EDTA.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting, LLC
40864 Arkansas Ave
Bennett, CO 80102
Phone: 303-644-4538
Fax: 720-859-4110
pruegg <@t> ihctech.net

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth
Chlipala
Sent: Friday, November 09, 2012 11:00 AM
To: 'Rene J Buesa'; Diana McCaig; 'histonet <@t> lists.utsouthwestern.edu'
Subject: RE: [Histonet] decal affecting IHC

Diana

I agree with Jack, formic acid is the way to go in my opinion, we use 10%
formic acid routinely but have pushed it up to 20% on occasion when time was
an issue.  HCL can work if its controlled really well but you don't have
much wiggle room, you can over decal quite quickly, formic acid is a bit
more forgiving.  As for Rene's comment we have seen the exact opposite with
respects to the EDTA decal part.  Granted we do not run a lot of IHC on EDTA
decaled samples but on several occasions and with several antibodies we were
able to obtain good IHC staining in formic acid decaled samples but those
antibodies did not work well in EDTA decaled samples.  These were not direct
comparisons of the decalcification method on the same study, but on samples
from different studies so other factors could have affected the results.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Friday, November 09, 2012 10:47 AM
To: Diana McCaig; 'histonet <@t> lists.utsouthwestern.edu'
Subject: Re: [Histonet] decal affecting IHC

IHC reactions can be affected by decalcification especially if:
1- the tissue is not perfectly fixed before going into decalcification
2- decalcification is done at temperature above room temp.
3- decal solution is too acid, specially made with inorganic acids.
If at all possible decalcification should be done with a chelating agent,
like EDTA
René J.


________________________________
From: Diana McCaig <dmccaig <@t> ckha.on.ca>
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, November 9, 2012 12:34 PM
Subject: [Histonet] decal affecting IHC

Does decalcifying tissue  (Calex II) affect the antibody reaction for IHC in
bony tissue?

Diana

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