Re: [Histonet] specimen size for IHC
CM Van der Loos is the "Dr. House of Histology". I believe whatever he
I pulled this from the Jan 06 archives.
[Histonet] RE: optimal thickness for cutting of IHC sections
"C.M. van der Loos"
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
quantifying an IHC technique wasn't influenced by tissue thickness
due to microtome abberations. We did cut (cryostat) sections of
different thickness and guess what: there was hardly any influence
on the staining intensity. 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:
immunohistochemistry. The measurement of the ratio’s between
collagen types I and I II. Histochem J (1994) 26,
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center
NL-1105 AZ Amsterdam
The Nether lands
Date: Tue, 3 Jan 2006 13:09:20 -0500
Subject : RE:
[Histonet] optimal thickness for cutting of IHC sections< BR>To:
I believe section thickness is less critical for IHC
because antibodies are very large molecules that don't
penetrate tissue very well, so regardless of
the thickness of the section, you are really only staining
exposed surface of the tissue, perhaps to a depth of 2
microns or so. We have verified this by electron
microscopy. This is quite different from standard histochemical
procedures where a thicker section results in a
more intense stain because the small dye molecules penetrate
the tissue readily and stain it all the way through.
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05/18/2006 01:45 PM
[Histonet] specimen size for IHC
The optimum size for IHC is 3-4mm thick and 1-2cm square to allow adequate
penetration in the fixation process. The Immunochemical Staining
handbook(3rd Ed, published by Dako) states to cut at 4-6.
In your invaluable, but, no doubt differing opinions, what is the
optimal section thickness for formalin fixed paraffin processed
tissues for immunostaining, using DAB as chromogen?, there has been
some discussion here whether it should be 4,5,6, or 7microns, many
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