[Histonet] RE: Histonet Digest, Vol 128, Issue 17

[Jamal]

I totally agree with you in step 4, and I think the tissue was not fixed
probably or not processed in good solutions, my suggestion for the time now
problem is to stain the slides manual.


Best Regards,


Jamal M. Al Rowaihi	Anatomic Pathology Supervisor       | Al Borg
Medical Laboratories |  Mobile +966 503629832|
j.rowaihi <@t> alborglaboratories.com 
Palestine St, Al Rajhi Building, P.O. Box 52817, Jeddah 21573, KSA    |
Phone: +966 12 670 0099	      | Fax: +966 12 676 4984     |
www.alborglaboratories.com


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Riesen,
Rebecca
Sent: Saturday, July 19, 2014 6:41 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Histonet Digest, Vol 128, Issue 17

Liz had some great suggestions! The one that I have found to be the MOST
important is #4.  Standing my slides up at a slight angle and allowing the
moisture to drain out from under the section AND then letting the drained
slides sit and dry for a good long time is extremely important for adherance
to be successful (I've never gone as far as a week, although it couldn't
hurt, but at least overnight seems necessary).  I would focus on that, so
you first get good adherance and then the other tips following the proper
drying will then help keep them on.  Good luck!

------------------------------

Message: 4
Date: Fri, 18 Jul 2014 14:44:13 +0000
From: "Murphy, Valerie" <murphyv <@t> karmanos.org>
Subject: [Histonet] Brain tissue coming off slide
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
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<A0CD799E6FCD4949AAD4A25377936712AE1DADC2 <@t> EX-MB2.kci-net.karmanos.org>
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We are staining human FFPE brain sections for Her 2 Neu and we're having a
problem with tissue coming off the slide during the HIER step.
We are currently using Vectabond coated plus slides and using a decloaking
chamber for HIER.
Are there any other steps we can take to prevent  tissue coming off the
slide?

Thank you,

Valerie Ratliff
Karmanos Cancer Institute


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Message: 5
Date: Fri, 18 Jul 2014 09:16:04 -0600
From: Elizabeth Chlipala <liz <@t> premierlab.com>
Subject: [Histonet] RE: Brain tissue coming off slide
To: "Murphy, Valerie" <murphyv <@t> karmanos.org>,
	"'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<14E2C6176416974295479C64A11CB9AE019C79ECD373 <@t> SBS2K8.premierlab.local>
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Valerie

There are a few things that you can try.

1.  This may or may not work with the Her2 IHC, since for that antibody I
believe it requires retrieval at least at 98C but time and temperature are
related with respects to heat retrieval.  You could try 70C for 2 hours this
is gentler on the tissue and works for some antibodies but not all of them.
For example it does not work for Ki-67 in our hands.  We use this retrieval
method on bone and other types of friable tissue if necessary.  

2.  We have found that some samples after HIER may require manual staining
methods in order to optimize tissue adherence, not sure if that is an option
for you too.

3.  Post fixation in 10% NBF after deparaffinization for about 10% minutes.
This may help tissue adherence but we have also found that this may decrease
the sensitivity of the protocol and you may have to adjust your primary
antibody concentration.

4.  Tissue adherence to the slide especially with brain (we have seen this
on rodent brains) requires proper fixation and adequate processing, poorly
fixed or processed brain samples will have issues with tissue adherence.

5.  Allow the cut slides to sit for about a week prior to staining we find
this may help with tissue adherence.

Good Luck

Liz

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

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