RE: [Histonet] Xylene Substitute for Counterstain Clearing
Andrew There are aqueous permanent mounting medias such as Advantage Permanent Mounting Media from Accurate Chemical NB300A (516) 333-2221 its been years since I used it but it does work on some applications. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrew Coleman Sent: Friday, May 11, 2012 10:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Xylene Substitute for Counterstain Clearing 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
RE: [Histonet] Xylene Substitute for Counterstain Clearing
Andrew You could use Clearium from Leica. Clearium can either be coverslipped from xylene or isopropyl alcohol. Drying time from isopropyl is longer then xylene. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory Manager X-Cell Laboratories 716-250-9235 etx. 232 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrew Coleman Sent: Friday, May 11, 2012 12:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Xylene Substitute for Counterstain Clearing 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
Re: [Histonet] Xylene Substitute for Counterstain Clearing
I think that it is called Crystal Mount - but apply to section, allow to harden dip slide in clearing media and coverslip. I know that there must be others out there as well. Good Luck On Fri, May 11, 2012 at 12:25 PM, Andrew Coleman andrewcoleman...@gmail.com wrote: 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
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
RE: [Histonet] Xylene Substitute for Counterstain Clearing
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
