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

From:"C.M. van der Loos"


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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