reply: Acid Alcohol Fixation & Plastics?

From:Gayle Callis <>

Always an option. 

Question:  what kind of acid alcohol fixation?? Carnoys, acetic acid
formalin, need specifics please.  Acids are not too kind to plastics, in my
experience, you would have to remove acids totally so as to not interfere
with any plastic polymerization. Extra alcohol rinses just to get rid of acid.

Also, what about trying just alcohol fixation?  For classic PMMA embedded
bone metabolic diagnostic work, 70% ethanol was originally used to fix
bone. This may not be the correct concentration of alcohol in your
acid/alcohol fixative. Calculate % of alcohol in the acid/alcohol mixture,
it could be 90%??? or so, and leave out the acid. You may get results for
IHC without acid being involved. Your morphology will not be as good as
formalin fixed, but it may be adequate for immunostaining. 

The joy of methyl methacrylate is you can totally remove it from microtomed
sections and do immunostaining.  Check out Neil Hands publication, J of
Histotechnology approx 3 years back on doing antigen retrieval.  He used
formalin fixation AND he will be giving a workshop at NSH in Sept on
plastic work, etc.  He used several changes of warm xylene, retrieved
stringently with pressure cooker.    Neil has a huge panel of antibodies
that worked on PMMA embedded soft tissues using these technics.  

At 11:31 AM 7/20/01 -0500, you wrote:
>Hello HistoNet!	
>We are looking to bring plastics to our facility as a large part of our 
>research involves bone and other hard tissues. The HistoNet has been 
>exceptionally helpful thus far in providing information and references for 
>this type of work.  Thank you!  We have an upcoming project where 
>immunohistochemistry will be performed using an antibody where the vendor 
>suggests acid alcohol fixation.  We need to plastic embed the 
>tissue.  Based on my research to date, formalin seems to be the fixative of 
>choice for plastic work but, previous work done using this antibody on 
>formalin fixed tissue yielded negative results.  Is acid alcohol an option 
>for tissue to be plastic (methyl methacrylate) embedded?
>Thanks in advance for any help.
>Tracey Couse
>Petit Institute for Bioengineering & Biosciences
>Georgia Institute of Technology
>Atlanta, GA
Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology
Montana State Universi

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