Re: LCM and antigen retrieval

From:richard hazelton

Dear Denise,

My experience with LCM and IHC is that they are incompatible. RNA is
degraded during the immunostaining and DNA would also be denatured during
the antigen retrieval process. If you use frozen sections both the
antibodies should given good staining without AR, but the RNA wil still
suffer after the prolonged incubation of a double staining procedure. The
DNA may survive. The use of charged slides will make microdissection
difficult. Poly-L-Lysine is the only adhesive I found which will allow
microdissection, but this depends on section thickness, type of tissue etc.
CD68 in paraffin sections may be better demonstrated after enzyme pre
treatment rather than AR, however CD34 is sensitive to some enzymes, so you
may need to stain for it first.
I am interested to learn how others have "gotten this to work".
Good Luck !

-----Original Message-----
From: Denise Bland-Piontek 
Date: Wednesday, 1 August 2001 3:10
Subject: LCM and antigen retrieval

>Is anyone out there doing Laser Capture Microdissection on
>Immunoperoxidase stains that require antigen retrieval? We are using a
>double stain procedure with CD-68 and CD-34 (DAKO products), marking
>with DAB and Fast Red. We are using antigen retrieval and charged
>slides. We realize that charged slides are not recommended for LCM,
>however we do not want to loose the tissue during antigen retrieval.
>Others have somehow gotten this to work and I was wondering if anyone
>has any tricks they can share with me. Thanks in advance,
>Denise Bland-Piontek, HTL

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