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
   qu= antifying an IHC technique wasn't influenced by tissue thickness
   due  to=  microtome  abberations.  We did cut (cryostat) sections of
   different thi= ckness and guess what: there was hardly any influence
   on the staining i= ntensity. The only thing we observed was
   that   thicker  sections=  showed  a  higher  non-specific  background
   staining   than  thinner  sections.=  See:
  
CM  van der Lo   os,   MMH   Marijianowski   and   AE   Becker:  Quantification  in
   immunohistoche=  mistry.  The  measurement  of the ratio’s between
   collagen   types   I   and   I=   II.   Histochem   J  (1994)  26,
   347-354<=    /P>   
Chris  van der   Loos, PhD
Dept. of Pathology
Academic Medical Center
   M=  2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The
   Nether=    lands= 
   
 
   
Date:  Tue,  3  Jan 2006 13:09:20 -0500 
From:
   "Mo= nfils, Paul"
   <PMonfils@Lifespan.org>
Subject=   :  RE:
   [Histonet]  optimal  thickness  for  cutting  of  IHC sections<   BR>To: 'histonet@lists.utsouthwestern.edu'
=  
I  believe  section  thickness  is less critical for IHC
   because  anti=  bodies  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
   exposed
surface  of  the  tissue,  perhaps  = to a depth of 2
   microns  or so.  We have
verified this b= y electron
   microscopy.   This  is  quite  different  from standard<   BR>histochemical  procedures  where  a  thicker section results in a
   more  i= ntense
stain because the small dye molecules penetrate
   the    tissue    =   readily   and   stain
it   all   the   way
   through.

= 
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