[Histonet] Question on epithelial staining in plastic resin

[Gayle Callis]

Neil Hand did pressure cooker retrieval and published in J of 
Histotechnology, NM Hand, Chruch RJ.  Superheating using pressure cooking: 
its use and application in unmasking antigens embedded in methyl 
methacrylate. J Histotechnology, 21:231-236, 1988.   If you are a member of 
NSH, they will send a copy of this to you - not sure if it will cost you 
anything.  NSH website has application for doing this, or call them.

He also wrote a chapter on the subject in Gamble and Bancroft Theory and 
Practice of Histological Techniques, 6th Edition, 2007  chapter 29.

He removed MMA from thin tissue sections, 2 um or so, with 37C xylene,  10 
to 20 minutes, followed by a routine rehydration to distilled water.  He had 
great success with his IHC on MMA embedded tissues, with a panel of 200 
antibodies or so.  I am sure the thickness of the section will affect MMA 
removal, so time in the warm xylene may need to be extended and maybe an 
extra change to ensure plastic removal.

I am not sure if he has pdf's of his publications, but you can contact him 
at Hand Neil (Histopathology):   Neil.Hand <@t> nuh.nhs.uk

He is regarded as one of the experts for doing MMA/IHC work.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT 59715


----- Original Message ----- 
From: "Herrick, James L." <Herrick.James <@t> mayo.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, May 15, 2008 3:58 PM
Subject: [Histonet] Question on epithelial staining in plastic resin


Hello everyone,

Does anyone have experience staining for epithelial cells (pig arteries)
in MMA embedded specimens? So far I have run immuno for VWF, eNOS and am
currently trying a lectin stain using Griffonia (no success). I have
previously run immuno on this same tissue type for actin which looked
great.

I am using a steamer for antigen retrieval with the solution Retrievit-8
from InnoGenex. I have tried anywhere from 15 minutes to 1 hour in the
steamer. For deplastification I am using a Xylene, 2-MEA, Acetone and
ETOH protocol. I am thinking that this may be too harsh.

Any feedback on this issue would be greatly appreciated. Thanks for your
help.

Jim
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