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Author Topic: Wanting to use IHC on frozen tissue  (Read 4043 times)

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Offline TRobertson

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Wanting to use IHC on frozen tissue
« on: September 03, 2013, 11:09:10 PM »
I have some brain tissue and I was wanting to use IHC to examine the aromatase enzyme distribution, but all the tissue I currently have is frozen at -80C.  Is it possible to do IHC on this this tissue or do I need to collect new brains from my experiment?  If I can do IHC on frozen tissue are there any special considerations that I should incorporate into my protocol?

Thanks!

Wanting to use IHC on frozen tissue
« on: September 03, 2013, 11:09:10 PM »

Offline cstan

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Re: Wanting to use IHC on frozen tissue
« Reply #1 on: September 04, 2013, 10:43:05 AM »
Hello,

Frozen sections are best kept at -80C until needed so you should be good to go. The sections just need to be warmed for a few minutes before fixing in acetone. Because of the absence of formalin based fixative it eliminates the need for antigen retrieval step. But if the tissue has been fixed in formalin, you might want to consider an antigen retrieval step although because the frozen sections are "friable" in nature, it might make it difficult.

Here's a link to an article that discusses application of antigen retrieval on frozen sections as reference in case it applies to you:

Yamashita S, Okada Y. Application of heat induced antigen retrieval to aldehyde fixed fresh frozen sections. J Histochem Cytochem. 2005 Nov;53(11):1421-32.

Hope this helps!
« Last Edit: September 04, 2013, 11:01:40 PM by cstan »

Offline iLOVEihc

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Re: Wanting to use IHC on frozen tissue
« Reply #2 on: October 07, 2013, 05:35:16 AM »
don't bother with antigen retrieval as this will damage the tissues. Remove the tissue from the freezer and allow it to dry. Then fix in cold acetone from the freezer for 10 min. Allow the tissue to dry then place into TBS or whatever buffer you are using to wash, then proceed with the protocol as with FFPE IHC. don't do peroxidase block in methanol.  :)
IHClovER

Offline WWang1991

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Re: Wanting to use IHC on frozen tissue
« Reply #3 on: March 11, 2015, 03:24:40 AM »
You may need help from professional experts.  http://www.creative-bioarray.com/Services/Immunohistochemistry-IHC.htm
this site has combined academic and industrial experiences in IHC service, and you can ask them for help or any consultation.
The secret to happiness is to keep setting yourself new challenges.
http://blog.creative-bioarray.com/

Re: Wanting to use IHC on frozen tissue
« Reply #3 on: March 11, 2015, 03:24:40 AM »