[Histonet] Xylene Substitute for Counterstain Clearing

[gayle callis]

You are also  likely to have spheres dissolve in toluene based mounting
media.   Aqueous media may be the answer unless the neutral red runs out of
the tissue in the aqueous environment.   Hard to get the best of both worlds
if dye runs in aqueous media and spheres dissolve in solvent based .   Good
luck! 

Gayle Callis     

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Friday, May 11, 2012 12:23 PM
To: histonet <@t> lists.utsouthwestern.edu; Andrew Coleman
Subject: Re: [Histonet] Xylene Substitute for Counterstain Clearing

After staining dry the sections in an oven at 60ºC for 10 minutes. When
completely dried, coverslip as usual. Beware of the mounting medium solvent
because it may contain xylene as well. Use one mounting medium without
xylene.
René J.

--- On Fri, 5/11/12, Andrew Coleman <andrewcoleman131 <@t> gmail.com> wrote:


From: Andrew Coleman <andrewcoleman131 <@t> gmail.com>
Subject: [Histonet] Xylene Substitute for Counterstain Clearing
To: histonet <@t> lists.utsouthwestern.edu
Date: Friday, May 11, 2012, 12:25 PM


Hi all,

We are performing a neutral red counterstain on tissue sections containing
colored polystyrene microspheres. The spheres are inert to alcohol, but are
washed out when we clear with xylene to coverslip.
The spheres are also supposedly soluble in DMF, acetone, acetonitrile,
chloroform and methylene chloride for what its worth.

Is it reasonable to coverslip these slides in permanent mount without
clearing with xylene after dehydrating the tissue? Or does anyone know of a
substitute clearing agent with chemical properties dissimilar enough from
xylene that might be worth trying instead?

Thanks,

Andrew

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