[Histonet] sections falling off slides

[Liz Chlipala]

Robin

We run quite a few IHC's on goat bone femurs.  We use a very good plus
slide. Dry flat at 40C overnight and let them sit a few days before we
stain, we do not place them in a 60C oven since we find that the
articular cartilage may flip.  We have good success with proteinase K
with staining on the dako stainer for several antibodies.  If we have to
run HIER we use a 70C retrieval for 2 hours and then may choose to
manually stain depending upon the samples rather then placing on the
dako autostainer.  We have done this type of staining on porcine femurs
on 2x3 slides manually.  

I really think tissue adherence is dependent upon good fixation, optimal
decalcification and proper processing, if samples have been
insufficiently decaled or processed you are going to run into problems
with tissue adherence.  I can't comment to the lack of staining
intensity we always pilot a few slides prior to staining the entire
study just to make sure our methods will work for the particular tissues
we are currently working on.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robin
Dean
Sent: Monday, May 16, 2011 12:07 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] sections falling off slides

Hi All,

 

We are trying to do IHC for multiple antigens on goat knees. The
sections
are decalcified FFPE sections and we are using a Dako autostainer. We
are
currently trying air-drying for  1-2 days  and baking of tissue slides
before use in IHC..

The one previous time we tried baking, we noticed a decrease in IHC
signal.
The sections are pretty large, we tried cutting them down, but can't
decrease the size much.  We also  decreased the number of washes and
size of
washes on the Daok, which helped only slightly.  Does anyone have any
suggestions?

 

Thank you in advance.

 

Robin

 

Robin R. Dean, Ph.D.

Senior Scientist & Study Director

Comparative Biosciences, Inc.

786 Lucerne Dr.

Sunnyvale, CA

(408) 738-8060

robin_dean <@t> compbio.com

 

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