[Histonet] RE: pfa vs. formalin

Andrea T. Hooper anh2006 <@t> med.cornell.edu
Mon Aug 7 10:36:17 CDT 2006

I have found that for many mouse antigens, the presence of this 
methanol in 10% NBF is deleterious when doing fixed frozen 
immunohistochemistry (a necessary evil for when we work with muscle 
and bone). So when doing fixed frozen IHC I always make up 4% PFA in 
PBS from a 16% PFA ampoule. Works very well.

When fixing specimens for paraffin IHC we always use 10% NBF with no problems.

>The only difference is that commercial formaldehyde solution contains a
>stabilizer, usually 10 to 15% methanol.  Therefore, 10% NBF made up from
>such a solution contains 1 to 1.5% methanol, while 10% buffered NBP contains
>nothing but water, formaldehyde, and buffer salts.  For routine
>morphological studies this makes no difference whatsoever.  But for some
>special studies the presence of a small amount of methanol could be
>deleterious, and for any such procedures paraformaldehyde would be
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