[Histonet] RE: Brain tissue coming off slide

Elizabeth Chlipala liz <@t> premierlab.com
Fri Jul 18 10:16:04 CDT 2014


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


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

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Murphy, Valerie
Sent: Friday, July 18, 2014 8:44 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Brain tissue coming off slide

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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