RE: Frozen IHC

From:"Barry, Lilith" <>

After sectioning and air drying I usually store the slides in slide boxes,
well sealed in -80c freezer. 
For the immuno, I take the slides out of the freezer, air dry, then fix with
Acetone (in my case acetone:alc 70%:30%). 
I DON'T air dry before the immuno, but move fast into buffer. This is
because I have found that the morphology is much better preserved this way.
By the way, I work primarily with neuronal tissue.
If you have more questions please let me know.

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

Lilith Ohannessian-Barry                               
National Research Council                          
Institute of Biological Sciences                  

-----Original Message-----
From: Jim Manavis []
Sent: Tuesday, September 26, 2000 11:13 PM
To: Histonet
Subject: Frozen IHC

Can someone tell me their experience with frozen sections which are post
fixed in cold acetone prior to applying immuno histochemistry.
Which procedure do you find is superior?  Does it also vary depending on the
Ab used?
Post fix immediately, dry, leave them say, overnight in the -20 freezer, and
then apply IHC?
Do you air dry after sectioning, store overnight, then post fix just prior
to IHC?
Do you section, post fix and then apply IHC on the same day?

Thanks, Kathy Cash

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