[Histonet] IHC on fresh frozen

[Patti Loykasek]

After hearing a presentation by Sharon Lear describing some low temp antigen
retrieval that she did, we changed our method for frozen sections. We fix
the frozen sections in 10% nuetral buffered formalin for 30'-60', rinse,
then do a gentle pretreatment. The gentle pretreatment is usually 10mM
citrate buffer pH6 at 70 degrees C for 30'. Slides are cooled, and usual IHC
done. This has worked well for us. Our staining with this method is more
reliable & intense than with previous methods.

Patti Loykasek



> Hi, all
> 
> Recently I did some IHC(chromagen methods) on mouse fresh frozen
> tissues, mainly using insulin antibody on pancreas. The image is
> much fuzzier compare to paraffin embedding tissue. And the
> staining also smeared to acinar cells which surround the islet.
> 
> I airdried slide(>30min) and used a general acetone method(-20C
> 5min) to fix the tissue before I did IHC.
> 
> How can I get a relatively sharp staining on the fresh frozen
> tissue?Does anyone here have any experience on it? Any
> suggestions?
> 
> Many thanks and have a nice day,
> 
> Ann
> 
> 
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 



This e-mail message, including any attachments, is for the sole use of the 
intended recipients and may contain privileged information. Any unauthorized 
review, use, disclosure or distribution is prohibited. If you are not the intended 
recipient, please contact the sender by e-mail and destroy all copies of the 
original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. 
at (206) 374-9000.