Re: Subject: Frozen IHC

From:Kim Kusser <>


I cut sections, then leave them to dry at room temp (2hrs to 2days - I haven't noticed a difference in the quality of the sections). When I go to stain the sections, I bring them to room temp (if they've been in the freezer) then I fix at 4 degree C (in the fridge) acetone for 10 minutes...let dry for 10 minutes....then put in PBS for at least 5 minutes to remove OCT before I go onto my blocking steps.

As far as when you do IHC....I've done IHC on sections that have been at -70 and -20 for months, or at room temp 10 minutes. It works for me. But, as with anybodies IHC protocols...just because it works for them, doesn't mean it will work for you : )



Date: 26 Sep 2000 22:46:12 -0500
From: "Jim Manavis" <>
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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