Hi Doc.,
    Often you can fix staining by altering either the titration or the
incubation time. Using a prediluted CD3 from DAKO we stain for 20 minuets in
primary then 10 minuets in secondary and tertiary (LINK2 and LABEL2 from DAKO).
We have also gotten acceptable results with concentrated CD3 @ 1:400 with a 30
minute primary incubation time.
    The main point is that often you need to try a variety of titers and
incubations. Also excessive decalcification can be detrimental to staining
intensity so to rule this out as a possibility try using a tonsil for a control
as it does not need to be decalcified.
Amos Brooks

"Nader, Alexander" wrote:

> we tried both the polyklonal CD3 from DAko and the monoklonal CD3 from
> Novocastra. Both work well, but in my opinion, the monoclonal antibody
> doesn't stain all the T-cells, or at least, stains fewer lymphocytes than
> the DAKO polyklonal. HIER is done for 10 min in pH 6 citrate-buffer in a
> preasure cooker, dedection kit is from DAKO. All material is fixed in
> neutral buffered formalin for 24 hours, BM-trephines are decalcified for 24
> hours (at least) in EDTA on a shaking board (that reduces the time for
> complete decalcification a lot).
>
> Dr. Alexander Nader
> Path. Institut Hanuschkrankenhaus
> A 1140 Wien, Oesterreich
>
> Alexander.Nader@wgkk.sozvers.at <mailto:Alexander.Nader@wgkk.sozvers.at>
>
> privat: Alexander.Nader@univie.ac.at
> <mailto:Alexander.Nader@univie.ac.at>
>
> > -----Ursprüngliche Nachricht-----
> > Von: Richard Cartun [mailto:Rcartun@harthosp.org]
> > Gesendet am: Donnerstag, 06. Jänner 2000 16:35
> > An: Histonet
> > Betreff: CD3
> >
> > I am interested in hearing what Histoneters are using for the
> > detection of CD3+ cells in paraffin sections (pAb vs. mAb,
> > digestion vs. HIER, etc.).  Thanks.
> >
> > R. Cartun
> >
> >