[Histonet] Davidson's fixative and IHC question

Julio Benavides j.benavides at eae.csic.es
Wed Feb 24 03:37:53 CST 2016


Hi,

in my experience, Bouin´s fixative can sometimes give you problems  
with IHC. Both because it seriously interfere with some antigens (in  
my case, I was doing IHC for lentivirus and herpesvirus and more than  
24h of fixation would drastically reduce IHC labelling) but also  
because it gives you non-specific labelling (in may case, ovine  
neutrophils came up positive with that lentivurs Ac).

Julio



Robert Byrd via Histonet <histonet at lists.utsouthwestern.edu> escribió:

> Bouin's solution is an excellent fixative for fetal brains, we were still
> using it routinely when I trained 10 years ago but has (and already had in
> most places) largely fallen out of use due to the explosive risk associated
> with dried picric acid residue in drainpipes etc. It does work remarkably
> well on soft and macerated fetal brains though. I use 10% NBF now out of
> necessity but might be able to track down a Bouin's protocol if you are
> interested.
>
> On Tue, Feb 23, 2016 at 5:11 PM, Hawkins, Hal K. via Histonet <
> histonet at lists.utsouthwestern.edu> wrote:
>
>>
>> We briefly experimented with using a fixative for stillborn fetal brains
>> that was a mixture of equal volumes of 20% formalin and 95% ethanol.  The
>> gross preservation and firmness of these very soft brains was improved, and
>> the histology results were fine, but as one might predict, some antigens
>> were not preserved for immunohistochemistry.
>>
>> ________________________________________
>> From: Jeff Jurczak via Histonet [histonet at lists.utsouthwestern.edu]
>> Sent: Tuesday, February 23, 2016 3:45 PM
>> To: Bob Richmond
>> Cc: Histonet at lists.utsouthwestern.edu
>> Subject: Re: [Histonet] Davidson's fixative and IHC question
>>
>> Is this a good fixative for fetal brain? Anybody have any suggestions as
>> to protocols? Do any of the readers here do fetal (human) brain on a
>> regular basis?
>>
>> ----- Original Message -----
>>
>> From: "Bob Richmond via Histonet" <histonet at lists.utsouthwestern.edu>
>> To: "Histonet at lists.utsouthwestern.edu" <histonet at lists.utsouthwestern.edu
>> >
>> Sent: Tuesday, February 23, 2016 3:22:50 PM
>> Subject: Re: [Histonet] Davidson's fixative and IHC question
>>
>> Several queries about the possible effect of Davidson's fixative on
>> immunohistochemistry.
>>
>> I have a lot of experience with Davidson's fixative, but none with its
>> effect on IHC.
>>
>> Davidson's fixative is easily prepared:
>>
>> 3 parts tap water
>> 3 parts ethanol or reagent alcohol
>> 2 parts 37% formalin (not neutral buffered formalin)
>> 1 part glacial acetic acid
>>
>> It is quite destructive to H & E staining if fixation is at all prolonged.
>> Tissue should be taken out of it in not more much more than 24 hours.
>>
>> The history of Davidson's fixative is somewhat obscure - I have a good bit
>> of information on it. John Kiernan has said he'll work on ascertaining more
>> about its history if he ever has the time.
>>
>> Bob Richmond
>> Samurai Pathologist
>> Maryville TN
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