Re: Lung Embedding Protocol

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From:Mick Rentsch <ausbio@nex.com.au> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Dear all,
Immunologists have used tween 20 in their buffers and solutions for EIA and
ELISA for years as a surfactant without any supposed ill effects. It would
be probable Triton X-100 would do the same, but just to play it safe, why
not try Tween 20 first (Do not use Tween 80 as this lyses some cells)
Regards Mike(Downunder)
-----Original Message-----
From: Barry Rittman <brittman@mail.db.uth.tmc.edu>
To: histonet@pathology.swmed.edu <histonet@pathology.swmed.edu>
Date: Thursday, 21 January 1999 9:39
Subject: Re: Lung Embedding Protocol


>Katie,
>           there have been several approaches to processing lung and
>ensuring that the reagents are not excluded by the air in the bronchial
>tree.
>One approach was to use fixatives with low surface tension such as alcohol
>containing fixatives or to add  a small amount of surfactant such as
>triton X 100 to the fixative.
>I would agree with Sharon that a controlled positive pressure helps in the
>process but would recommend that you also include 0.01% -0.1% surfactant
>in the fixative. This has been shown to decrease surface tension without
>compromising routine histology. I am not sure if this has been used with
>IHC but would be interested to hear some comments about this.
>Another approach is to gently vacuum the fixative before use and use
>gentle vacuum to remove the air, once  in the fixative.
>I think that it is also advisable to vacuum infiltrate during paraffin
>processing.
>Barry
>
>Katie B wrote:
>
>> ---sbledsoe <sbledsoe@iupui.edu> wrote:
>> >
>> > I have been asked to Post the following:
>> >
>> > Could someone please suggest a good protocol for embedding lung
>> tissue in
>> > paraffin.
>> >
>
>




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