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G'day Sean,

You indicated the Drosophila tissue bits float on top of the fixative
without triton? In which case it is difficult to see how the fixative will
be able to have much of an effect on anything. I'd be sticking with the
Triton.

Steve
Stephen H. Cody

On 5 January 2011 12:07, Sean Speese <[log in to unmask]> wrote:

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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:
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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
> >
>