[Histonet] RE: optimal thickness for cutting of IHC sections

From:"C.M. van der Loos"

   Sorry  guys  my  message yesterday was somewhat distorted. Here it is   again...

   Dear Paul,

   I totally agree what you wrote about IHC: it's just a surface event.

   Long  time  ago  we  had  to  prove  that our (prehistoric) attempt of
   quantify=  ing  an IHC technique wasn't influenced by tissue thickness
   due  to  microtome=  aberrations.  We  did  cut (cryostat) sections of
   different  thickness  and gue= ss what: there was hardly any influence
   on  the  staining  intensity.  The  only=  thing  we observed was that
   thicker  sections  showed  a higher non-specific b= ackground staining
   than thinner sections. See:

   CM  van  der  Loos, MMH Marijianowski and AE Becker: Quantification in
   immu=  nohistochemistry.  The  measurement  of  the  ratio’s between
   collagen types I= and III. Histochem J (1994) 26, 347-354

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


   Date: Tue, 3 Jan 2006 13:09:20 -0500
   From: "Monfils, Paul" 
   Subject:  RE:  [Histonet]  optimal  thickness  for  cutti=  ng  of IHC
   To: [1]'histonet@lists.utsouthwestern.edu'   I  believe  section thickness is less critical for IHC because antibo   dies are
   very  large  molecules  that  don't  penetrate  tissue very well, so    regardless of
   the  thickness  of  the  section,  you  are  really only staining= the
   surface  of  the  tissue, perhaps to a depth of 2 microns or s= o.  We
   verified  this by electron microscopy.  This is qui= te different from
   histochemical  procedures  where a thicker sect= ion results in a more
   stain  because  the small dye molecules pene= trate the tissue readily
   and stain
   it all the way through.




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