[Histonet] IHC after EDTA decalcifying

[Houston, Ronald]

Perhaps I'm missing something here too. Did you decalcify the tonsil for the same length of time as the mouse femur? If you are looking at  e.g. CD markers in bone or indeed bone marrow, it is important that you have positive control material that you know works with undecalcified tissue and a corresponding piece of tissue that is treated in the same way as your test sample (i.e. 2 tonsil samples from the same case, 1 of which has been decalcified and the other not, in the same control block - we also have bone marrow control blocks that we do the same with)

It is too much of a generalization that EDTA preserves all antigen sites. Some are affected much more than others. 
So to be able to help at all, what specific markers are you looking at, what are they raised in, and how are you detecting the antibody-antigen reaction?

and I completely agree with what Ray says also

Ronnie

Ronnie Houston, MS HT(ASCP)QIHC FRMS
Anatomic Pathology Manager
ChildLab, a Division of Nationwide Children's Hospital
www.childlab.com

700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
(F) 614-722-2899
ronald.houston <@t> nationwidechildrens.org
www.NationwideChildrens.org



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of koellingr <@t> comcast.net
Sent: Thursday, August 15, 2013 10:17 PM
To: Colleen Forster
Cc: Histonet
Subject: Re: [Histonet] IHC after EDTA decalcifying



Could I ask exactly what you are looking for in mouse femur and human tonsil the tonsil  I assume is a control?  For what?  Whatever you are looking for is obviously in human tonsil.  Are you positive it is in murine femur.  I used to stain for 15-20 targets in EDTA decaled mouse femurs but my control was also a mouse. 



Ray 

Research Scientist 

Comparative Medicine 

University of Washington 


----- Original Message -----


From: "Colleen Forster" <cforster <@t> umn.edu>
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, August 15, 2013 4:58:35 PM
Subject: [Histonet] IHC after EDTA decalcifying 

Hello fellow histonetters, 

Can someone who has worked with decalcifyied bone tell me if EDTA interferes with IHC staining? I was under the impression it did not but cannot get staining in mouse femurs that I have decaled in EDTA. I have used both the steamer and the decloaker for retrieval and in the human tonsil the stain is great. 

Any suggestions...... 

Thanks in advance! 

Colleen Forster
U of MN 

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