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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, May 11, 2012 12:23 PM To: histonet@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 <andrewcoleman...@gmail.com> wrote: From: Andrew Coleman <andrewcoleman...@gmail.com> Subject: [Histonet] Xylene Substitute for Counterstain Clearing To: histonet@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet