Fixation times for Immuno methods.

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From:Victoria Baker <>
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Hi Everyone!

The controversy over fixation and IHC has reared its'
ugly little head again.

So rather than bang my head against the wall from
frustration, I thought I'd put it out and see what
others are doing.

The tissue is either rat or mouse, the size of the
tissue is usually 5-6mm in diameter and approximately
4mm in depth.  This is a research facility where time
isn't always critical (except when something is needed

So if anyone is working with this under similar
circumstances do you think you could take a minute and
just answer a couple of questions?  

1.  How long do you fix your tissues for? 10 hours,
>10 hours, 12 to 24 hours.

2.  Do you do fixation on or off of your processor?

3.  Do you use different fixatives for different

4.  How long is your overall processing time and what
do you use to hold tissue in after fixation?

5.  This one may cause some controversy.....Does
anyone cap their blocks after they have finished with
them for storage.  

6.  Does anyone use a xylene substitute and which one
do you use.

Thanks.  I've come to the conclusion that the humorous
"element" I sent out last night also pertains to
stubborn Principle investigators. Even the nicest of

Vikki Baker
American Health Foundation
Valhalla, New York

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