[Histonet] Bouin's without the picric acid?

[Geoff McAuliffe]

Hi Tyrone:

Picric acid may be bad for some silver stains but not for all. Williams 
fix is recommended for the Bodain method and I have used it with 
excellent results on mouse brain.
Also overnight immersion fixation of a whole fish - you did not say how 
large- is not going to give good results. The 3 components of Bouin all 
penetrate and fix at different rates.
I would fix for a week, room temp. with the head cut off and maybe the 
mandible removed to allow better penetration.
Bouin without picric acid is just an alcoholic formalin but if you want 
to avoid formalin (PFA) then how about an acid alchohol fix, 70 or 80 
percent EtOH with 5% acetic acid?

Geoff

On 9/14/2011 7:52 AM, Tyrone Genade wrote:
> Hello,
>
> I have been using Bouin's fixative on my fish specimens. I had simply
> been killing the fish, slitting them down their belly and letting them
> lie in Bouin's overnight. This worked well to soften the tissue and in
> particular the brain case (I'm interested in the brain but the
> dissection is delicate and I'm clumsy). After trying some silver
> staining the results were not very good. I have since learned that
> picric acid interferes with silver staining (and fluorescent staining
> like FluoroJade-B) so no more picric acid for me.
>
> My question is this: can I use Buoin's without picric acid? The
> solution is 75 mL saturated picric acid, 25 mL 40% formaldehyde and 5
> mL glacial acetic acid to make 100 mL.  If I simply left out the
> picric acid, replacing it with distilled water, would this cause
> trouble? I am reluctant to switch to PFA as this can destroy certain
> antibody epitopes whereas the Bouin's doesn't (so I'm told but I was
> also told that Buoin's won't cause trouble at all...).
>
> My other idea was to use PFA and then decalcify in dilute acetic or formic acid.
>
> Any suggestions or advice?
>
> Thanks


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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
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