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January 2011

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Confocal Microscopy List <[log in to unmask]>
Subject:
Re: Question about surfactants to break surface tension
From:
Sean Speese <[log in to unmask]>
Date:
Tue, 4 Jan 2011 17:07:43 -0800
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Hi Carl,
   Point taken regarding the exterior staining, thanks.  My fear is that you
are right about the properties of surfactants.

Sean

On Tue, Jan 4, 2011 at 4:59 PM, Boswell, Carl A - (cboswell) <
[log in to unmask]> wrote:

> Hi Sean,
> Don't know of an alternative that can do what you want.  I would think that
> the same properties of a surfactant that make it one would also disturb
> lipid layers.
>
> Aside from that, I would caution against the assumption that fixation
> without detergent allows only exterior staining.  The fixation process will
> also open holes in membrane and allow Ab to penetrate.  At least test it
> before committing to that route.  The best result I've had restricting
> immunostaining to a surface is to stain live material, then fix after
> washing.
> C
>
> Carl A. Boswell, Ph.D.
> Molecular and Cellular Biology
> Univ. of Arizona
> 520-954-7053
> FAX 520-621-3709
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]]
> On Behalf Of Sean Speese
> Sent: Tuesday, January 04, 2011 5:44 PM
> To: [log in to unmask]
> Subject: Question about surfactants to break surface tension
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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>
> Hi all,
>   Bit of an off topic question, but we do lots of immunos on small bits of
> Drosophila tissue that always float on top of the fix due to the surface
> tension.  We can use low amounts of triton (.01%) to overcome this, but I
> would like to be able to leave detergents out until I am convinced the
> fixation is good.  In some cases I would like to do staining of only things
> on the cell surface, and thus need to leave detergents completely out of the
> staining procedure.  Therefore, does anyone know of a good surfactant that
> would break the surface tension but not permeabilize cells?
>
> Thanks,
>   Sean Speese
>
>

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