RE: IHC on frozen muscle

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From:Louise Taylor <>
To:histonet <>
Content-Type:text/plain; charset="iso-8859-1"

Hi Sharon, Donald & Bonnie who have thus far answered my mail

Regarding the section adherence of my sections  - the normal muscle controls
stuck beautifully, its just the dystrophy ones - I guess it could have been
a bad batch of silane slides, but I'm still looking for answers.....


-----Original Message-----
From: sharon lang []
Sent: 09 June 2000 02:02
Subject: Re: IHC on frozen muscle

I would try the charged slides, especially with frozen sectioning.
Take care

>From: Louise Taylor <>
>To: histonet <>
>Subject: IHC on frozen muscle
>Date: Thu, 08 Jun 2000 15:31:31 +0200
>Hi histonetters,
>I am doing dystrophy markers (dystrophin, sarcoglycan etc) on cryostat
>sections of  fresh frozen muscle. These biosies have been frozen in
>isopentane cooled in liquid nitrogen. I have noted that there is a lot of
>fat in these biosies(part of the pathology?), and that section adherence
>through the staining process is poor ( non existent actually) even though I
>am using silane treated slides.
>  I was wondering if anyone has had a similar problem, and what
>countermeasures could be taken. I though that maybe fixation in acetone
>prior to IHC might improve matters, but I don't know whether this will
>impact negatively on the staining, I have already optimised my panel of 12
>markers  on "unfixed" muscle treated the same way, and would not like to
>have to reoptimise.
>  Any thoughts? I would appreciate some help here, as muscle frozens are
>quite my usual sample.
>best regards
>louise taylor
>Research Laboratory
>Department of Anatomical Pathology
>South African Institute for Medical Research
>South Africa

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