[Histonet] RE: optimal thickness for cutting of IHC sections
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
sections
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
exposed
surface of the tissue, perhaps to a depth of 2 microns or s= o. We
have
verified this by electron microscopy. This is qui= te different from
standard
histochemical procedures where a thicker sect= ion results in a more
intense
stain because the small dye molecules pene= trate the tissue readily
and stain
it all the way through.
=
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References
1. 3D"javascript:main.compose('new','t='hi_______________________________________________
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