[Histonet] RE: antigen retrieval for IHC

[C.M. van der Loos]

   Dear Patsy and others,

   Recently  we did  something  similar  with a tonsil that was cut in 10
   pieces  and  formalin-fixed from 12 hours up to one year. Per fixation
   time  three  samples  were  taken for a tissue array. For example, our
   staining  results  showed that at least caspase-3 nicely stained the 1
   year-fixed  tissue  sample.  Ki67-staining  "died"  after  3 months of
   fixation.

   Chris van der Loos, PhD
   Dept. of Pathology
   Academic Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   phone:  +31 20 5665631
   fax:    +31 20 6960389


   Date: Mon, 26 Sep 2005 09:58:46 -0600
   From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
   Subject: RE: antigen retrieval for IHC

               Shi  mentions  in  his  book  on pg.10 ANTIGEN RESTORATION
   "Simply
   bathing  deparaffinized sections in a cold 20% sucrose-saline solution
   could,
   over several days, restore a certain amount of immunoreactivity."
   In  Introduction  to  IHC by Polak and Van Noorden on pg 24 3.8.1 "The
   simplest
   form  of  reversing the effects of formalin is to wash the tissue well
   before
   processing"
   I am about to test these statements, as I just received tissues for an
   IHC
   project  that  have  been  in 10% NBF for over one year.  I washed the
   tissues
   in  running tap h20 for 2 hrs., I will now process them into paraffin.
   I am
   planning as well to put some of the sections in 20% sucrose at 4dc for
   2
   days  if  I have trouble getting good IHC signal.  I will keep you all
   posted
   on  this  experiment.   These  are samples of mouse mammory tissue.  I
   will be
   testing them with cleaved caspase 3 ! and Ki67