[Histonet] IHC in resin embedded sections

From:"Tessa Murray"

   Dear Histonetters,

   I  am  looking  for  some advice about the feasibility of doing IHC on
   tissues embedded for EM work.  I need to section very small tissues in
   a  specific orientation which is difficult to determine when embedding
   in  paraffin.   I  was  wondering  if we could borrow the EM technique
   where  (as  I  understand)  the  tissue  is embedded in resin and then
   temporarily  fixed to a "peg" for sectioning - thus allowing us to cut
   some  sections,  check  the  orientation  and  then  rotate  the block
   accordingly  until  we get the orientation exactly right.  Obviously I
   am  concerned  about  how  well  antibodies are going to recognize the
   tissue  after  processing, how are the sections de-resined, is there a
   specific  antigen  retreival  step  for  resin  tissues etc.  Any help
   appreciated  -  perhaps  there is a way to adjust sectioning angles in
   FFPE  tissues  that  I've  not  thought of?  Frozen tissues are not an
   option  as we want to preserve morphology as much as possible.  Cheers

   Tessa J Murray PhD
   Tufts University School of Medicine
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